Modulators of the eIF2alpha pathway

ABSTRACT

Provided herein, inter alia, are compounds and methods useful for modulating the translational effects of eIF2α phosphorylation, the Integrated Stress Response (ISR), and the unfolded protein response (UPR); for treating diseases; for increasing protein production, and for improving long-term memory.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/849,428, filed Sep. 9, 2015, which is a continuation of InternationalPatent Application No. PCT/US2014/029568, filed Mar. 14, 2014, whichclaims the benefit of U.S. Provisional Patent Application No.61/787,633, filed Mar. 15, 2013, all of which are incorporated herein byreference in their entirety and for all purposes.

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED AS AN ASCII TEXT FILE

The Sequence Listing written in file 84850-903323_ST25.TXT, created Mar.14, 2014, 3,576 bytes, machine format IBM-PC, MS-Windows operatingsystem, is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

In metazoa, diverse stress signals converge at a single phosphorylationevent at serine 51 of a common effector, the translation initiationfactor eIF2α. This step is carried out by four eIF2α kinases inmammalian cells: PERK, which responds to an accumulation of unfoldedproteins in the endoplasmic reticulum (ER), GCN2 to amino acidstarvation and UV light, PKR to viral infection, and HRI to hemedeficiency. This collection of signaling pathways has been termed the“integrated stress response” (ISR), as they converge on the samemolecular event. eIF2α phosphorylation results in an attenuation oftranslation with consequences that allow cells to cope with the variedstresses (1).

eIF2 (which is comprised of three subunits, α, β and γ) binds GTP andthe initiator Met-tRNA to form the ternary complex(eIF2-GTP-Met-tRNA_(i)), which, in turn, associates with the 40Sribosomal subunit scanning the 5′UTR of mRNAs to select the initiatingAUG codon. Upon phosphorylation of its α-subunit, eIF2 becomes acompetitive inhibitor of its GTP-exchange factor (GEF), eIF2B (2). Thetight and nonproductive binding of phosphorylated eIF2 to eIF2B preventsloading of the eIF2 complex with GTP thus blocking ternary complexformation and reducing translation initiation (3). Because eIF2B is lessabundant than eIF2, phosphorylation of only a small fraction of thetotal eIF2 has a dramatic impact on eIF2B activity in cells.

Paradoxically, under conditions of reduced protein synthesis, a smallgroup of mRNAs that contain upstream open reading frames (uORFs) intheir 5′UTR are translationally upregulated (4,5). These includemammalian ATF4 (a cAMP element binding (CREB) transcription factor) andCHOP (a pro-apoptotic transcription factor) (6-8). ATF4 regulates theexpression of many genes involved in metabolism and nutrient uptake andadditional transcription factors, such as CHOP, which is under bothtranslational and transcriptional control (9). Phosphorylation of eIF2αthus leads to preferential translation of key regulatory molecules anddirects diverse changes in the transcriptome of cells upon cellularstress.

One of the eIF2α kinases, PERK, lies at the intersection of the ISR andthe unfolded protein response (UPR) that maintains homeostasis ofprotein folding in the ER (10). The UPR is activated by unfolded ormisfolded proteins that accumulate in the ER lumen because of animbalance between protein folding load and protein folding capacity, acondition known as “ER stress”. In mammals, the UPR is comprised ofthree signaling branches mediated by ER-localized transmembrane sensors,PERK, IRE1, and ATF6. These sensor proteins detect the accumulation ofunfolded protein in the ER and transmit the information across the ERmembrane, initiating unique signaling pathways that converge in theactivation of an extensive transcriptional response, which ultimatelyresults in ER expansion (11). The lumenal stress-sensing domains of PERKand IRE1 are homologous and likely activated in analogous ways by directbinding to unfolded peptides (12). This binding event leads tooligomerization and trans-autophosphorylation of their cytosolic kinasedomains, and, for PERK, phosphorylation of its only known substrate,eIF2α. In this way, PERK activation results in a quick reduction in theload of newly synthesized proteins that are translocated into theER-lumen (13).

Upon ER stress, both the transcription factor XBP1s, produced as theconsequence of a non-conventional mRNA splicing reaction initiated byIRE1, and the transcription factor ATF6, produced by proteolysis andrelease from the ER membrane, collaborate with ATF4 to induce the vastUPR transcriptional response. Transcriptional targets of the UPR includethe ER protein folding machinery, the ER-associated degradationmachinery, and many other components functioning in the secretorypathway (14). Although the UPR initially mitigates ER stress and as suchconfers cytoprotection, persistent and severe ER stress leads toactivation of apoptosis that eliminates damaged cells (15,16).

Small-molecule therapeutics that inhibit the UPR and/or the IntegratedStress Response could be used in cancer as a single agent or incombination with other chemotherapeutics [1] [2] [3], for enhancement oflong-term memory [5] [6], in neurodegenerative and prion associateddiseases [4], in white matter disease (VWM) [7] and in biotechnologyapplications that would benefit from increased protein translation.

Disclosed herein, inter alia, are solutions to these and other problemsin the art.

BRIEF SUMMARY OF THE INVENTION

In a first aspect is provided a method of treating an integrated stressresponse-associated disease in a patient in need of such treatment, themethod including administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, to the patient,wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In another aspect is provided a method of treating a disease associatedwith phosphorylation of eIF2α in a patient in need of such treatment,the method including administering a therapeutically effective amount ofa compound, or a pharmaceutically acceptable salt thereof, to thepatient, wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In another aspect is provided a method of treating a disease in apatient in need of such treatment, the method including administering atherapeutically effective amount of a compound to the patient, whereinthe disease is selected from the group consisting of cancer, aneurodegenerative disease, vanishing white matter disease, childhoodataxia with CNS hypo-myelination, and an intellectual disabilitysyndrome; and wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In another aspect is provided a method of treating an inflammatorydisease in a patient in need of such treatment, the method includingadministering a therapeutically effective amount of a compound, or apharmaceutically acceptable salt thereof, to the patient, wherein thecompound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In another aspect is provided a method of improving long-term memory ina patient, the method including administering a therapeuticallyeffective amount of a compound to the patient, wherein the compound is acompound described herein.

In another aspect is provided a method of increasing protein expressionof a cell or in vitro expression system, the method includingadministering an effective amount of a compound to the cell orexpression system, wherein the compound is a compound described herein.

In another aspect is provided a compound, or a pharmaceuticallyacceptable salt thereof, having the formula:

wherein ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, z4, z5and z6, are as described herein, including embodiments and in the methodof treatment section herein above. In embodiments, Ring A is substitutedor unsubstituted cycloalkylene or substituted or unsubstituted arylene.In embodiments, L¹, L², L³, and L⁴ are independently a bond, —NH—, —O—,—S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene. In embodiments, R¹, R³, R⁵,R⁶ and R⁷ are independently hydrogen, halogen, —OCH₃, —OCH₂Ph, —C(O)Ph,—CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂,—C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃, —C(NH—NH)CF₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R² and R⁴ areindependently ═NR⁷, =0, or ═S. The symbols z2 and z4 are eachindependently 0 or 1. The symbols z5 and z6 are each independently aninteger from 0 to 5.

In another aspect is provided a pharmaceutical composition including apharmaceutically acceptable excipient and a compound, orpharmaceutically acceptable salt thereof, as described herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1C. High-throughput cell-based screen for inhibitors of PERKsignaling; (FIG. 1A) schematic representation of the ATF4 luciferasereporter used in the primary screen; the 5′ UTR of human ATF4 containingthe uORFs 1 and 2 was fused to firefly luciferase and inserted into aretroviral expression system; (FIG. 1B) primary screen optimization;HEK293T stably expressing the ATF4 luciferase reporter were plated in384 well plates and treated for 6 h with 100 nM thapsigargin (Tg) orDMSO as a no ER stress control; luciferase production was measured atthe end point after 6 h; the Z′ was calculated as 1−(3(σTg−σDMSO)/(μg−μDMSO)); (FIG. 1C) primary screen results; the ATF4luciferase reporter cell line was treated for 6 h with 100 nMthapsigargin and library compounds (10 μM); inhibition of the luciferaseactivity reporter was calculated as the percent reduction in relativeluminescence normalized to thapsigargin treatment (0% inhibition) andthe no-ER stress control (100% inhibition); compounds were consideredhits if they lied beyond 3 standard deviations (SD) from thethapsigargin treatment mean (line).

FIGS. 2A-2D. Identification of ISRIB as a potent cell-based inhibitor ofPERK signaling; (FIG. 2A) structures of ISRIB isosteromers; (FIG. 2B)inhibition of the ATF4 luciferase reporter in HEK293T cells by ISRIBstereoisomers; inhibition is plotted in relation to the concentration ofeither the cis or trans isomer of ISRIB; cells were treated with 2 μg/mlof tunicamycin to induce ER stress and different concentrations of theinhibitors for 7 h (N=2); (FIG. 2C) effect of ISRIB on production ofendogenous ATF4, PERK phosphorylation, and XBP1s production; animmunoblot analysis of PERK, ATF4 and XBP1s in HEK293T cells treatedwith different ER stress inducers (2.5 μg/ml tunicamycin (Tm) or 100 nMthapsigargin (Tg)) with or without 200 nM ISRIB for 3 h is shown; thearrowhead marks the XBP1s specific band; (FIG. 2D) effect of ISRIB onXBP1 mRNA splicing; TAQMAN® assays for XBPlunspliced (XBP1u) and XBP1spliced (XBP1s) on cDNA synthesized from total RNA extracted from U2OScells treated with 2 μg/ml of tunicamycin in the presence or absence of200 nM ISRIB for the indicated times are shown; percent splicing wascalculated as the ratio of XBP1 s over total XBP1 mRNA (XBP1u+XBP1s).

FIGS. 3A-3F. ISRIB makes cells resistant to eIF2α phosphorylation; (FIG.3A) ISRIB does not block eIF2α phosphorylation upon ER stress; eIF2αphosphorylation was measured using an Alpha-Screen Surefire eIF2αp-S51assay; U2OS cells were plated in 96 well plates and treated with 2 μg/mltunicamycin or 100 nM thapsigargin in the presence or absence of 100 nMISRIB for the indicated times or with ISRIB alone for 120 m (N=4, SD);see FIG. 7A for supporting Western blot analysis of eIF2αphosphorylation; (FIG. 3B) ISRIB blocks global translational attenuationobserved after eIF2α phosphorylation during ER stress; HEK293T cellswere treated with 100 nM thapsigargin and 200 nM ISRIB for either 1 or 3h prior to a 20 min pulse with ³⁵S methionine before lysis; equalamounts of lysate were loaded on an SDS-PAGE gel and quantification ofradiolabeled methionine incorporation of lysates was done by geldensitometry (N=2, SD) using Image J; (see FIG. 7B for SDS-PAGE); (FIG.3C) polysome gradient analysis showing the block in global translationalattenuation upon addition of ISRIB on ER-stressed cells; MEFs were grownin the presence or absence of 2 μg/ml of tunicamycin with or without 200nM ISRIB for 1 h; cell lysates were loaded on a 10-50% sucrose gradient,centrifuged at 150,000×g for 2.4 h and absorbance at 254 nm was measuredacross the gradient (see FIG. 7C for quantitation of polysome profile);a representative experiment is shown (N=3); (FIG. 3D) cells treated withISRIB are resistant to the global translational attenuation exerted byforced expression of eIF2α(S51D); HEK293Trex cells were transduced witha tetracycline inducible phospho-mimetic (S51D) allele of eIF2α;transgene expression was induced by addition of 25 nM doxycycline for 14h in the presence or absence of 200 nM ISRIB; lysates were collected andanalyzed as described in (FIG. 3C above (see FIG. 7D for quantitation ofpolysome profile); a representative experiment is shown (N=2); (FIG. 3E)ISRIB does not reverse global translational attenuation exerted throughinhibition of CAP-dependent initiation; wild-type MEFs were treated with750 nM torin-1 in the presence or absence of 200 nM ISRIB for 2 h;lysates were collected and analyzed as described in FIG. 3C above; arepresentative experiment is shown (N=2); (FIG. 3F) ISRIB blocksproduction of ATF4 upon GCN2 or HRI activation; an immunoblot analysisof PERK, ATF4 and total eIF2α in HEK293T cells starved for cysteine andmethionine or treated with an HRI activator (6 μM) for 5 h in thepresence or absence of 200 nM ISRIB is shown; tunicamycin was used as apositive control for induction of ATF4 and the shift in PERK mobility;under amino acid starvation we consistently observe a partial reductionof ATF4 production by ISRIB by Western blot analysis but observe acomplete block in induction of the ATF4 luciferase reporter (see FIG.7E).

FIGS. 4A-4B. ISRIB impairs induction of the transcriptional networkcontrolled by ATF4; (FIG. 4A) ER-stress dependent induction of CHOP andGADD34 mRNA is impaired in cells treated with ISRIB; qPCR analysis oftotal RNA extracted from U2OS cells treated with 2 μg/ml of tunicamycinin the presence or absence of 200 nM ISRIB for the indicated times; mRNAlevels for each sample were normalized to GAPDH (N=4); P values arederived from a one tail Student's t-test for unpaired samples;statistical significance: CHOP, P=0.0006 (*); GADD34, P=0.0008 (*);(FIG. 4B) ISRIB blocks CHOP production during ER stress; animmunofluorescence analysis of U2OS cells treated with 100 nMthapsigargin for 2 h in the presence or absence of 200 nM ISRIB isshown; a secondary Alexa Dye 488 anti-mouse antibody andrhodamine-phalloidin were used to visualize CHOP and actin,respectively.

FIGS. 5A-5D. ISRIB impairs adaptation to ER-stress prolonging activationof the UPR sensors; (FIG. 5A) ISRIB sensitizes cells to acute ER stress;HEK293T cells were subjected with an acute dose of tunicamycin (2μg/mL), ISRIB (200 nM) or a combination of both for 24 h; the treatedcells were equally diluted to a concentration that would allow singlecell clonal expansion and re-seeded onto 6-well plates in a 3-folddilution series; clonal colonies were visualized by Crystal Violetstain; (FIG. 5B) ISRIB synergizes with ER stress to activate caspase3/7; Hela cells were plated in 96 well plates and treated with 5 μg/mlof tunicamycin or 500 nM thapsigargin with or without 25 nM ISRIB forthe indicated times; caspase3/7 activation was measured usingCELLPLAYER™ kinetic caspase 3/7 reagent and cells were imaged in anINCUCYTE™ system; green object count/mm² representing caspace-3/7activation was measured at 2 h intervals (See FIG. 8A for endpointquantitation of % cells with activated caspase 3/7); (FIG. 5C) IRE1oligomers are sustained on ER-stressed cells treated with ISRIB;confocal microscopy micrographs of HEK293Trex cells carrying aninducible GFP-tagged IRE1 allele were treated with 10 nM doxycycline for24 h to induce the transgene, followed by treatment with 5 μg/ml oftunicamycin in the presence or absence of 200 nM ISRIB for the indicatedtimes; (See FIG. 8B for corresponding XBP1 mRNA splicing data); (FIG.5D) ATF6 cleavage is sustained in ER-stressed cells treated with ISRIB;immunoblot analysis of ATF6 processing in HEK293Trex cells carrying aninducible FLAG epitope-tagged ATF6; cells were treated with 50 nMdoxycycline for 18 h to induce the transgene followed by treatment with100 nM thapsigargin in the presence or absence of 200 nM ISRIB for theindicated times; total eIF2α is used as a loading control.

FIG. 6A-6E. ISRIB enhances spatial and fear-associated learning inrodents; (FIG. 6A) escape latencies are significantly shorter in micetreated with ISRIB. Data (means+/−SEM) were obtained in a weak 5days-long training session in the hidden platform version of the Morriswater maze (1 trial per day); mean escape latencies were plotted as afunction of training days in mice treated with ISRIB (closed squares,N=8) or vehicle (open circles N=8) (P<0.05, (*)); mice were injecteddaily with ISRIB immediately after training; (FIG. 6B) after completionof training in the study shown in FIG. 6A above, mice treated with ISRIB(black column) showed a significant preference for the target quadrant(P<0.05, (*)); the probe test was performed 24 h after the last trainingsession; P values are derived from a two-tailed Student's t test forunpaired samples; (FIG. 6C) after completion of training in the studyshown in FIG. 6A above, mice treated with ISRIB (black column) increasedthe number of times they crossed the platform location as compared tothe vehicle-treated mice (grey column) (P<0.05, (*)); P values arederived from a two-tailed Student's t test for unpaired samples; (FIG.6D) chronic systemic administration of ISRIB (intraperitoneally for 4consecutive days) enhances long-term contextual fear memory (right bars,24 h), while it does not affect short-term memory (left bars, 1 h) (n=8per group, p<0.05, (*)); data are presented as mean±SEM; (FIG. 6E)auditory fear conditioning is enhanced in rats treated with ISRIB;freezing in response to a tone was assessed 3 h (short-term memory, STM,left panel) and 24 h (long-term memory, LTM, right panel) after training(vehicle-treated N=8, and ISRIB-treated N=7) after tone presentation(CS) and before tone presentation (pre-CS); for these experimentsvehicle or ISRIB was infused directly by cannula into the amygdala aftertraining; ISRIB-infused rats show increase freezing at 24 h (P<0.05,(*)).

FIG. 7A-7E. ISRIB makes cells resistant to eIF2α phosphorylation; (FIG.7A) ISRIB does not inhibit eIF2α phosphorylation; immunoblot analysis ofPERK, ATF4, phospho-eIF2α and total eIF2α in HEK293T cells treated withor without 2 μg/ml of tunicamycin or 100 nM thapsigargin in the presenceor absence of 200 nM ISRIB for 3 h; (FIG. 7B) ISRIB blocks translationalattenuation upon ER stress; autoradiogram (top) and total protein(bottom) obtained from HEK293T cells that were treated with 100 nMthapsigargin with or without 200 nM ISRIB for either 1 or 3 h prior to a20 min pulse with ³⁵S methionine before lysis; equal amounts of lysatewere loaded on an SDS-PAGE gel; (FIG. 7C) ISRIB blocks translationalattenuation upon ER stress; the polysome profile in FIG. 3C wasquantitated by calculating the area under the curve corresponding to themonosome peak (80S), or the area under the curve corresponding to thetrace covering the polysome region and then plotted as a ratio over thearea under the curve corresponding to the peak of the 60S subunit; (FIG.7D) ISRIB sustains translation upon expression of eIF2α(S51D); thepolysome profile in FIG. 3D was quantitated as described in FIG. 7C;(FIG. 7E) ISRIB blocks induction of the ATF4 luciferase translationalreporter upon HRI and GCN2 activation; HEK293T carrying the ATF4luciferase reporter were treated with 2 μg/ml of tunicamycin to induceER stress, 6 μM of the HRI activator or grown in media lacking cysteineand methionine for 7 h in the presence or absence of 200 nM ISRIB (N=4);the relative luciferase units are normalized to the no treatmentcontrol; using this reporter we observe a smaller fold change inproduction of luciferase by amino acid starvation that is fully blockedby addition of ISRIB.

FIGS. 8A-8C. ISRIB impairs adaptation to ER-stress prolonging activationof the UPR sensors; (FIG. 8A) ISRIB synergizes with ER-stress to inducecaspase 3/7; green object count/mm² representing caspace-3/7 activationdepicted in FIG. 5A was normalized to the total number of cells at twodifferent endpoints; in order to quantify the total number of cells,VYBRANT® DYECYCLE™ Green staining solution (1 μM) was added directly tothe well immediately after the Caspase-3/7 scan and incubated for 1 hprior to acquiring final images at both 46 and 72 h; data is presentedas % cells with activated caspase 3/7 at these two endpoints; note thatby 72 h the ER-stress inducing conditions used in this experiment are sodetrimental that they diminish the synergistic effects observed byaddition of ISRIB; the synergy was clearly seen at the 46 h time-point;(FIG. 8B) XBP1 splicing is sustained in ER-stressed cells upon additionof ISRIB; HEK293T cells were treated with tunicamycin (2 μg/ml) for theindicated times in the presence or absence of 200 nM ISRIB; (FIG. 8C)RNA was isolated from the cells and reverse transcribed followed by PCRwith oligos that amplify both the unspliced and spliced versions of XBP1mRNA or GAPDH; the DNA was electrophoresed in a 2.5% agarose gel; theasterix (*) denotes a hybrid PCR product.

FIG. 9. Overview of SMDC 750213/ISRIB analogs.

FIG. 10. Overview of unfolded protein response.

FIG. 11. Overview of integrated stress response.

FIG. 12. Overview of regulation of ATF4 translation.

FIG. 13. Overview of PERK cell based screen.

FIG. 14. ISRIB as a potent cell based inhibitor of the PERK branch.

FIG. 15. ISRIB blocks the translational attenuation induced by ERstress.

FIG. 16. ISRIB makes cells resistant to eIF2α phosphorylation.

FIG. 17. ISRIB blocks the PERK branch of the UPR.

FIG. 18. ISRIB decreases viability of cells subjected to ER stress.

FIG. 19. Regulation of memory consolidation via eIF2α phosphorylation.

FIG. 20. ISRIB increases spatial learning.

FIG. 21. ISRIB increases auditory fear learning in rats.

FIG. 22. RMPI 8226 cells were implanted subcutaneously in BALB/c scidmice; at day 26, when tumors had an average size of 95 mm3, mice wereorally dosed (daily) with vehicle or 5 mg/kg of ISRIB; tumor size wasmeasured twice weekly and the mean tumor volume was plotted as afunction of study days (error bars show standard error of the mean);after 26 days of dosing, a significant difference in tumor size isobserved between ISRIB-treated and the vehicle-treated control group(non-parametric t-test Mann-Whitney p<0.0001)

FIGS. 23A-23B. ISRIB enhances translation in both ER-stressed (FIG. 23A)and unstressed (FIG. 23B) RPMI8226 cells; polysome gradient analysis ofRPMI cells in the presence or absence of 5 μg/ml of tunicamycin with orwithout 1 μM ISRIB; cell lysates were fractionated on a sucrosegradient; addition of ISRIB leads to a decrease in the 80S peak andincrease in the polysome population.

FIG. 24. ISRIB enhances production of luciferase in a rabbitreticulocyte in vitro translation assay; luciferase mRNA was added tothe lysates in the presence or absence of different concentrations ofISRIB; the amount of luciferase protein produced was quantitated byaddition of One-Glo and the Relative luminescent units were normalizedto no-addition control.

FIG. 25. Inflammation and proposed mechanism for postsurgical cognitivedysfunction.

FIG. 26. Experimental design for measuring postsurgical cognitivedysfunction and effects of ISRIB on cognitive function; measurement ofmemory is a behavioral test using trace fear conditioning (TFC), whichaims to establish a permanent memory in animals by using sensorialinformation; associative learning is presented with a neutralconditioning stimulus, paired with an aversive unconditioning stimulus(e.g. shock); freezing by the animal corresponds to the ability of theanimal to retain memory from the context in which it has been trained.Separate cohorts of animals were used to assess inflammatory status 24 hafter surgery. Downwards arrows represent injection points.

FIG. 27. Trace fear conditioning: hippocampal-dependent memory test;postsurgical cognitive dysfunction measured in animal model andmeasurement of effects of ISRIB on cognitive function; mice subjected tosurgery exhibited reduced freezing when compared to control mice atpostoperative day three; perioperative administration of ISRIB mitigatedmemory impairment after surgery. Control animals were given vehiclesolution.

FIG. 28. Inflammation measurements based on IL-6 serum levels as aninflammation marker; measuring the effects of ISRIB on systemicinflammation; mice subjected to surgery exhibited an increase in serumIL-6 24 h after surgery when compared to control mice; perioperativeadministration of ISRIB abrogated the increase in IL6. Levels weremeasured using an enzyme-linked immunosorbent kit.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The abbreviations used herein have their conventional meaning within thechemical and biological arts. The chemical structures and formulae setforth herein are constructed according to the standard rules of chemicalvalency known in the chemical arts.

Where substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents that would result from writing thestructure from right to left, e.g., —CH₂O— is equivalent to —OCH₂—.

The term “alkyl,” by itself or as part of another substituent, means,unless otherwise stated, a non-cyclic straight (i.e., unbranched) orbranched carbon chain (or carbon), or combination thereof, which may befully saturated, mono- or polyunsaturated and can include di- andmultivalent radicals, having the number of carbon atoms designated(i.e., C₁-C₁₀ means one to ten carbons). Examples of saturatedhydrocarbon radicals include, but are not limited to, groups such asmethyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl,sec-butyl, (cyclohexyl)methyl, homologs and isomers of, for example,n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkylgroup is one having one or more double bonds or triple bonds. Examplesof unsaturated alkyl groups include, but are not limited to, vinyl,2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and thehigher homologs and isomers. An alkoxy is an alkyl attached to theremainder of the molecule via an oxygen linker (—O—).

The term “alkylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkyl, asexemplified, but not limited by, —CH₂CH₂CH₂CH₂—. Typically, an alkyl (oralkylene) group will have from 1 to 24 carbon atoms, with those groupshaving 10 or fewer carbon atoms being preferred in the presentinvention. A “lower alkyl” or “lower alkylene” is a shorter chain alkylor alkylene group, generally having eight or fewer carbon atoms. Theterm “alkenylene,” by itself or as part of another substituent, means,unless otherwise stated, a divalent radical derived from an alkene.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a non-cyclic stable straight or branchedchain, or combinations thereof, including at least one carbon atom andat least one heteroatom selected from the group consisting of O, N, P,Si, and S, and wherein the nitrogen and sulfur atoms may optionally beoxidized, and the nitrogen heteroatom may optionally be quaternized. Theheteroatom(s) 0, N, P, S, and Si may be placed at any interior positionof the heteroalkyl group or at the position at which the alkyl group isattached to the remainder of the molecule. Examples include, but are notlimited to: —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃,—CH₂—S—CH₂—CH₃, —CH₂—CH 2, —S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃,—Si(CH₃)₃, —CH₂—CH═N—OCH₃, —CH═CH—N(CH₃)—CH₃, —O—CH₃, —O—CH₂—CH₃, and—CN. Up to two or three heteroatoms may be consecutive, such as, forexample, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃.

Similarly, the term “heteroalkylene,” by itself or as part of anothersubstituent, means, unless otherwise stated, a divalent radical derivedfrom heteroalkyl, as exemplified, but not limited by,—CH₂—CH₂—S—CH₂—CH₂— and —CH₂—S—CH₂—CH₂—NH—CH₂—. For heteroalkylenegroups, heteroatoms can also occupy either or both of the chain termini(e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, andthe like). Still further, for alkylene and heteroalkylene linkinggroups, no orientation of the linking group is implied by the directionin which the formula of the linking group is written. For example, theformula —C(O)₂R′— represents both —C(O)₂R′— and —R′C(O)₂—. As describedabove, heteroalkyl groups, as used herein, include those groups that areattached to the remainder of the molecule through a heteroatom, such as—C(O)R′, —C(O)NR′, —NR′R″, —OR′, —SR′, and/or —SO₂R′. Where“heteroalkyl” is recited, followed by recitations of specificheteroalkyl groups, such as —NR′R″ or the like, it will be understoodthat the terms heteroalkyl and —NR′R″ are not redundant or mutuallyexclusive. Rather, the specific heteroalkyl groups are recited to addclarity. Thus, the term “heteroalkyl” should not be interpreted hereinas excluding specific heteroalkyl groups, such as —NR′R″ or the like.

The terms “cycloalkyl” and “heterocycloalkyl,” by themselves or incombination with other terms, mean, unless otherwise stated,non-aromatic cyclic versions of “alkyl” and “heteroalkyl,” respectively,wherein the carbons making up the ring or rings do not necessarily needto be bonded to a hydrogen due to all carbon valencies participating inbonds with non-hydrogen atoms. Additionally, for heterocycloalkyl, aheteroatom can occupy the position at which the heterocycle is attachedto the remainder of the molecule. Examples of cycloalkyl include, butare not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples ofheterocycloalkyl include, but are not limited to,1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl, 2-piperazinyl, and the like. A “cycloalkylene” and a“heterocycloalkylene,” alone or as part of another substituent, means adivalent radical derived from a cycloalkyl and heterocycloalkyl,respectively.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” includes, but is not limited to, fluoromethyl,difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

The term “acyl” means, unless otherwise stated, —C(O)R where R is asubstituted or unsubstituted alkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

The term “aryl” means, unless otherwise stated, a polyunsaturated,aromatic, hydrocarbon substituent, which can be a single ring ormultiple rings (preferably from 1 to 3 rings) that are fused together(i.e., a fused ring aryl) or linked covalently. A fused ring aryl refersto multiple rings fused together wherein at least one of the fused ringsis an aryl ring. The term “heteroaryl” refers to aryl groups (or rings)that contain at least one heteroatom such as N, O, or S, wherein thenitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. Thus, the term “heteroaryl” includesfused ring heteroaryl groups (i.e., multiple rings fused togetherwherein at least one of the fused rings is a heteroaromatic ring). A5,6-fused ring heteroarylene refers to two rings fused together, whereinone ring has 5 members and the other ring has 6 members, and wherein atleast one ring is a heteroaryl ring. Likewise, a 6,6-fused ringheteroarylene refers to two rings fused together, wherein one ring has 6members and the other ring has 6 members, and wherein at least one ringis a heteroaryl ring. And a 6,5-fused ring heteroarylene refers to tworings fused together, wherein one ring has 6 members and the other ringhas 5 members, and wherein at least one ring is a heteroaryl ring. Aheteroaryl group can be attached to the remainder of the moleculethrough a carbon or heteroatom. Non-limiting examples of aryl andheteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl,1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl,4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl,5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl,4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl,2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl,5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl,5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and6-quinolyl. Substituents for each of the above noted aryl and heteroarylring systems are selected from the group of acceptable substituentsdescribed below. An “arylene” and a “heteroarylene,” alone or as part ofanother substituent, mean a divalent radical derived from an aryl andheteroaryl, respectively. Non-limiting examples of heteroaryl groupsinclude pyridinyl, pyrimidinyl, thiophenyl, thienyl, furanyl, indolyl,benzoxadiazolyl, benzodioxolyl, benzodioxanyl, thianaphthanyl,pyrrolopyridinyl, indazolyl, quinolinyl, quinoxalinyl, pyridopyrazinyl,quinazolinonyl, benzoisoxazolyl, imidazopyridinyl, benzofuranyl,benzothienyl, benzothiophenyl, phenyl, naphthyl, biphenyl, pyrrolyl,pyrazolyl, imidazolyl, pyrazinyl, oxazolyl, isoxazolyl, thiazolyl,furylthienyl, pyridyl, pyrimidyl, benzothiazolyl, purinyl,benzimidazolyl, isoquinolyl, thiadiazolyl, oxadiazolyl, pyrrolyl,diazolyl, triazolyl, tetrazolyl, benzothiadiazolyl, isothiazolyl,pyrazolopyrimidinyl, pyrrolopyrimidinyl, benzotriazolyl, benzoxazolyl,or quinolyl. The examples above may be substituted or unsubstituted anddivalent radicals of each heteroaryl example above are non-limitingexamples of heteroarylene.

A fused ring heterocyloalkyl-aryl is an aryl fused to aheterocycloalkyl. A fused ring heterocycloalkyl-heteroaryl is aheteroaryl fused to a heterocycloalkyl. A fused ringheterocycloalkyl-cycloalkyl is a heterocycloalkyl fused to a cycloalkyl.A fused ring heterocycloalkyl-heterocycloalkyl is a heterocycloalkylfused to another heterocycloalkyl. Fused ring heterocycloalkyl-aryl,fused ring heterocycloalkyl-heteroaryl, fused ringheterocycloalkyl-cycloalkyl, or fused ringheterocycloalkyl-heterocycloalkyl may each independently beunsubstituted or substituted with one or more of the substitutentsdescribed herein.

The term “oxo,” as used herein, means an oxygen that is double bonded toa carbon atom.

The term “alkylsulfonyl,” as used herein, means a moiety having theformula —S(O₂)—R′, where R′ is a substituted or unsubstituted alkylgroup as defined above. R′ may have a specified number of carbons (e.g.,“C₁-C₄ alkylsulfonyl”).

Each of the above terms (e.g., “alkyl,” “heteroalkyl,” “aryl,” and“heteroaryl”) includes both substituted and unsubstituted forms of theindicated radical. Preferred substituents for each type of radical areprovided below.

Substituents for the alkyl and heteroalkyl radicals (including thosegroups often referred to as alkylene, alkenyl, heteroalkylene,heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, andheterocycloalkenyl) can be one or more of a variety of groups selectedfrom, but not limited to, —OR′, ═O, ═NR′,

═N—OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —CONR′R″,—OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′,—NR—C(NR′R″R′″)═NR″″, —NR—C(NR′R″)═NR′″, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″,—NRSO₂R′, —NR′NR″R′″, —ONR′R″,

—NR′C═(O)NR″NR′″R″″, —CN, —NO₂, monophosphate (or derivatives thereof),diphosphate (or derivatives thereof), triphosphate (or derivativesthereof), in a number ranging from zero to (2 m′+1), where m′ is thetotal number of carbon atoms in such radical. R, R′, R″, R′″, and R″″each preferably independently refer to hydrogen, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl (e.g., aryl substituted with 1-3 halogens),substituted or unsubstituted heteroaryl, substituted or unsubstitutedalkyl, alkoxy, or thioalkoxy groups, or arylalkyl groups. When acompound of the invention includes more than one R group, for example,each of the R groups is independently selected as are each R′, R″, R′″,and R″″ group when more than one of these groups is present. When R′ andR″ are attached to the same nitrogen atom, they can be combined with thenitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example,—NR′R″ includes, but is not limited to, 1-pyrrolidinyl and4-morpholinyl. From the above discussion of substituents, one of skillin the art will understand that the term “alkyl” is meant to includegroups including carbon atoms bound to groups other than hydrogengroups, such as haloalkyl (e.g., —CF₃ and —CH₂CF₃) and acyl (e.g.,—C(O)CH₃, —C(O)CF₃, —C(O)CH₂OCH₃, and the like).

Similar to the substituents described for the alkyl radical,substituents for the aryl and heteroaryl groups are varied and areselected from, for example: —OR′, —NR′R″, —SR′, -halogen, —SiR′R″R′″,—OC(O)R′, —C(O)R′, —CONR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″,—NR″C(O)₂R′, —NR—C(NR′R″R′″)═NR″″, —NR—C(NR′ R″)═NR′″, —S(O)R′,—S(O)₂R′, —S(O)₂NR′R″, —NRSO₂R′, —NR′NR″R′″, —ONR′R″,NR′C═(O)NR″NR′″R″″, —CN, —NO₂, —R′, —N₃, —CH(Ph)₂, fluoro(C₁-C₄)alkoxy,and fluoro(C₁-C₄)alkyl, in a number ranging from zero to the totalnumber of open valences on the aromatic ring system; and where R′, R″,R′″, and R″″ are preferably independently selected from hydrogen,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, andsubstituted or unsubstituted heteroaryl. When a compound of theinvention includes more than one R group, for example, each of the Rgroups is independently selected as are each R′, R″, R′″, and R″″ groupswhen more than one of these groups is present.

Two or more substituents may optionally be joined to form aryl,heteroaryl, cycloalkyl, or heterocycloalkyl groups. Such so-calledring-forming substituents are typically, though not necessarily, foundattached to a cyclic base structure. In one embodiment, the ring-formingsubstituents are attached to adjacent members of the base structure. Forexample, two ring-forming substituents attached to adjacent members of acyclic base structure create a fused ring structure. In anotherembodiment, the ring-forming substituents are attached to a singlemember of the base structure. For example, two ring-forming substituentsattached to a single member of a cyclic base structure create aspirocyclic structure. In yet another embodiment, the ring-formingsubstituents are attached to non-adjacent members of the base structure.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally form a ring of the formula -T-C(O)—(CRR)_(q)—U—, whereinT and U are independently —NR—, —O—, —CRR′—, or a single bond, and q isan integer of from 0 to 3. Alternatively, two of the substituents onadjacent atoms of the aryl or heteroaryl ring may optionally be replacedwith a substituent of the formula -A-(CH₂)_(r)—B—, wherein A and B areindependently —CRR′—, —O—, —NR—, —S—, —S(O)—, —S(O)₂—, —S(O)₂NR′—, or asingle bond, and r is an integer of from 1 to 4. One of the single bondsof the new ring so formed may optionally be replaced with a double bond.Alternatively, two of the substituents on adjacent atoms of the aryl orheteroaryl ring may optionally be replaced with a substituent of theformula —(CRR′)_(s)—X′— (C″R″R′″)_(d)—, where s and d are independentlyintegers of from 0 to 3, and X′ is —O—, —S—, —S(O)—, —S(O)₂—, or—S(O)₂NR′—. The substituents R, R′, R″, and R′″ are preferablyindependently selected from hydrogen, substituted or unsubstitutedalkyl, substituted or unsubstituted heteroalkyl, substituted orunsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl,substituted or unsubstituted aryl, and substituted or unsubstitutedheteroaryl.

As used herein, the terms “heteroatom” or “ring heteroatom” are meant toinclude, oxygen (O), nitrogen (N), sulfur (S), phosphorus (P), andsilicon (Si).

A “substituent group,” as used herein, means a group selected from thefollowing moieties:

-   -   (A) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂,        —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,        —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH,        —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃, unsubstituted alkyl,        unsubstituted heteroalkyl, unsubstituted cycloalkyl,        unsubstituted heterocycloalkyl, unsubstituted aryl,        unsubstituted heteroaryl, and    -   (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,        heteroaryl, substituted with at least one substituent selected        from:        -   (i) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂,            —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,            —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH,            —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃, unsubstituted alkyl,            unsubstituted heteroalkyl, unsubstituted cycloalkyl,            unsubstituted heterocycloalkyl, unsubstituted aryl,            unsubstituted heteroaryl, and        -   (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,            heteroaryl, substituted with at least one substituent            selected from:            -   (a) oxo, halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂,                —NO₂, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,                —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H,                —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃,                unsubstituted alkyl, unsubstituted heteroalkyl,                unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl, and            -   (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl,                aryl, heteroaryl, substituted with at least one                substituent selected from: oxo,            -   halogen, —CF₃, —CN, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —SH,                —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,                —NHC═(O)NHNH₂, —NHC═(O) NH₂, —NHSO₂H, —NHC═(O)H,                —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, —NHSO₂CH₃, —N₃,                unsubstituted alkyl, unsubstituted heteroalkyl,                unsubstituted cycloalkyl, unsubstituted                heterocycloalkyl, unsubstituted aryl, unsubstituted                heteroaryl.

A “size-limited substituent” or “size-limited substituent group,” asused herein, means a group selected from all of the substituentsdescribed above for a “substituent group,” wherein each substituted orunsubstituted alkyl is a substituted or unsubstituted C₁-C₂₀ alkyl, eachsubstituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl.

A “lower substituent” or “lower substituent group,” as used herein,means a group selected from all of the substituents described above fora “substituent group,” wherein each substituted or unsubstituted alkylis a substituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl.

In some embodiments, each substituted group described in the compoundsherein is substituted with at least one substituent group. Morespecifically, in some embodiments, each substituted alkyl, substitutedheteroalkyl, substituted cycloalkyl, substituted heterocycloalkyl,substituted aryl, substituted heteroaryl, substituted alkylene,substituted heteroalkylene, substituted cycloalkylene, substitutedheterocycloalkylene, substituted arylene, and/or substitutedheteroarylene described in the compounds herein are substituted with atleast one substituent group. In other embodiments, at least one or allof these groups are substituted with at least one size-limitedsubstituent group. In other embodiments, at least one or all of thesegroups are substituted with at least one lower substituent group.

In other embodiments of the compounds herein, each substituted orunsubstituted alkyl may be a substituted or unsubstituted C₁-C₂₀ alkyl,each substituted or unsubstituted heteroalkyl is a substituted orunsubstituted 2 to 20 membered heteroalkyl, each substituted orunsubstituted cycloalkyl is a substituted or unsubstituted C₃-C₈cycloalkyl, each substituted or unsubstituted heterocycloalkyl is asubstituted or unsubstituted 3 to 8 membered heterocycloalkyl, eachsubstituted or unsubstituted aryl is a substituted or unsubstitutedC₆-C₁₀ aryl, and/or each substituted or unsubstituted heteroaryl is asubstituted or unsubstituted 5 to 10 membered heteroaryl. In someembodiments of the compounds herein, each substituted or unsubstitutedalkylene is a substituted or unsubstituted C₁-C₂₀ alkylene, eachsubstituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 20 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₈cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 8 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 10 membered heteroarylene.

In some embodiments, each substituted or unsubstituted alkyl is asubstituted or unsubstituted C₁-C₈ alkyl, each substituted orunsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8membered heteroalkyl, each substituted or unsubstituted cycloalkyl is asubstituted or unsubstituted C₃-C₇ cycloalkyl, each substituted orunsubstituted heterocycloalkyl is a substituted or unsubstituted 3 to 7membered heterocycloalkyl, each substituted or unsubstituted aryl is asubstituted or unsubstituted C₆-C₁₀ aryl, and/or each substituted orunsubstituted heteroaryl is a substituted or unsubstituted 5 to 9membered heteroaryl. In some embodiments, each substituted orunsubstituted alkylene is a substituted or unsubstituted C₁-C₈ alkylene,each substituted or unsubstituted heteroalkylene is a substituted orunsubstituted 2 to 8 membered heteroalkylene, each substituted orunsubstituted cycloalkylene is a substituted or unsubstituted C₃-C₇cycloalkylene, each substituted or unsubstituted heterocycloalkylene isa substituted or unsubstituted 3 to 7 membered heterocycloalkylene, eachsubstituted or unsubstituted arylene is a substituted or unsubstitutedC₆-C₁₀ arylene, and/or each substituted or unsubstituted heteroaryleneis a substituted or unsubstituted 5 to 9 membered heteroarylene. In someembodiments, the compound is a chemical species set forth in theExamples section below.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds that are prepared with relatively nontoxic acidsor bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, e.g., Berge et al., Journal of Pharmaceutical Science 66:1-19(1977)). Certain specific compounds of the present invention containboth basic and acidic functionalities that allow the compounds to beconverted into either base or acid addition salts. Otherpharmaceutically acceptable carriers known to those of skill in the artare suitable for the present invention. Salts tend to be more soluble inaqueous or other protonic solvents that are the corresponding free baseforms. In other cases, the preparation may be a lyophilized powder in 1mM-50 mM histidine, 0.1%-2% sucrose, 2%-7% mannitol at a pH range of 4.5to 5.5, that is combined with buffer prior to use.

Thus, the compounds of the present invention may exist as salts, such aswith pharmaceutically acceptable acids. The present invention includessuch salts. Examples of such salts include hydrochlorides,hydrobromides, sulfates, methanesulfonates, nitrates, maleates,acetates, citrates, fumarates, tartrates (e.g., (+)-tartrates,(−)-tartrates, or mixtures thereof including racemic mixtures),succinates, benzoates, and salts with amino acids such as glutamic acid.These salts may be prepared by methods known to those skilled in theart.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers from the various salt forms in certain physical properties, suchas solubility in polar solvents.

In addition to salt forms, the present invention provides compounds,which are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

As used herein, the term “salt” refers to acid or base salts of thecompounds used in the methods of the present invention. Illustrativeexamples of acceptable salts are mineral acid (hydrochloric acid,hydrobromic acid, phosphoric acid, and the like) salts, organic acid(acetic acid, propionic acid, glutamic acid, citric acid and the like)salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like)salts.

Certain compounds of the present invention possess asymmetric carbonatoms (optical or chiral centers) or double bonds; the enantiomers,racemates, diastereomers, tautomers, geometric isomers, stereoisometricforms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as (D)- or (L)- for amino acids, and individual isomers areencompassed within the scope of the present invention. The compounds ofthe present invention do not include those which are known in art to betoo unstable to synthesize and/or isolate. The present invention ismeant to include compounds in racemic and optically pure forms.Optically active (R)- and (S)-, or (D)- and (L)-isomers may be preparedusing chiral synthons or chiral reagents, or resolved using conventionaltechniques. When the compounds described herein contain olefinic bondsor other centers of geometric asymmetry, and unless specified otherwise,it is intended that the compounds include both E and Z geometricisomers.

As used herein, the term “isomers” refers to compounds having the samenumber and kind of atoms, and hence the same molecular weight, butdiffering in respect to the structural arrangement or configuration ofthe atoms.

The term “tautomer,” as used herein, refers to one of two or morestructural isomers which exist in equilibrium and which are readilyconverted from one isomeric form to another.

It will be apparent to one skilled in the art that certain compounds ofthis invention may exist in tautomeric forms, all such tautomeric formsof the compounds being within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude all stereochemical forms of the structure; i.e., the R and Sconfigurations for each asymmetric center. Therefore, singlestereochemical isomers as well as enantiomeric and diastereomericmixtures of the present compounds are within the scope of the invention.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of a hydrogen by a deuterium ortritium, or the replacement of a carbon by ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I), or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areencompassed within the scope of the present invention.

The symbol “

” denotes the point of attachment of a chemical moiety to the remainderof a molecule or chemical formula.

The terms “a” or “an,” as used in herein means one or more. In addition,the phrase “substituted with a[n],” as used herein, means the specifiedgroup may be substituted with one or more of any or all of the namedsubstituents. For example, where a group, such as an alkyl or heteroarylgroup, is “substituted with an unsubstituted C₁-C₂₀ alkyl, orunsubstituted 2 to 20 membered heteroalkyl,” the group may contain oneor more unsubstituted C₁-C₂₀ alkyls, and/or one or more unsubstituted 2to 20 membered heteroalkyls. Moreover, where a moiety is substitutedwith an R substituent, the group may be referred to as “R-substituted.”Where a moiety is R-substituted, the moiety is substituted with at leastone R substituent and each R substituent is optionally different.

Descriptions of compounds of the present invention are limited byprinciples of chemical bonding known to those skilled in the art.Accordingly, where a group may be substituted by one or more of a numberof substituents, such substitutions are selected so as to comply withprinciples of chemical bonding and to give compounds which are notinherently unstable and/or would be known to one of ordinary skill inthe art as likely to be unstable under ambient conditions, such asaqueous, neutral, and several known physiological conditions. Forexample, a heterocycloalkyl or heteroaryl is attached to the remainderof the molecule via a ring heteroatom in compliance with principles ofchemical bonding known to those skilled in the art thereby avoidinginherently unstable compounds.

The terms “treating” or “treatment” refers to any indicia of success inthe treatment or amelioration of an injury, disease, pathology orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,pathology or condition more tolerable to the patient; slowing in therate of degeneration or decline; making the final point of degenerationless debilitating; improving a patient's physical or mental well-being.The treatment or amelioration of symptoms can be based on objective orsubjective parameters; including the results of a physical examination,neuropsychiatric exams, and/or a psychiatric evaluation. For example,certain methods herein treat cancer (e.g. pancreatic cancer, breastcancer, multiple myeloma, cancers of secretory cells), neurodegenerativediseases, vanishing white matter disease, childhood ataxia with CNShypo-myelination, inflammatory diseases (e.g. postsurgical cognitivedysfunction or traumatic brain injury), and/or intellectual disabilitysyndromes (e.g. associated with impaired function of eIF2 or componentsin a signal transduction or signaling pathway including eIF2). Forexample certain methods herein treat cancer by decreasing or reducing orpreventing the occurrence, growth, metastasis, or progression of cancer;treat neurodegeneration by improving mental wellbeing, increasing mentalfunction, slowing the decrease of mental function, decreasing dementia,delaying the onset of dementia, improving cognitive skills, decreasingthe loss of cognitive skills, improving memory, decreasing thedegradation of memory, or extending survival; treat vanishing whitematter disease by reducing a symptom of vanishing white matter diseaseor reducing the loss of white matter or reducing the loss of myelin orincreasing the amount of myelin or increasing the amount of whitematter; treat childhood ataxia with CNS hypo-myelination by decreasing asymptom of childhood ataxia with CNS hypo-myelination or increasing thelevel of myelin or decreasing the loss of myelin; treat an intellectualdisability syndrome by decreasing a symptom of an intellectualdisability syndrome, treat cancer by decreasing a symptom of cancer,treat neurodegeneration by treating a symptom of neurodegeneration;treat an inflammatory disease (e.g. postsurgical cognitive dysfunctionor traumatic brain injury) by treating a symptom of the inflammatorydisease (e.g. postsurgical cognitive dysfunction or traumatic braininjury). Symptoms of cancer (e.g. pancreatic cancer, breast cancer,multiple myeloma, cancers of secretory cells), neurodegenerativediseases, vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2), or inflammatory diseases (e.g.postsurgical cognitive dysfunction or traumatic brain injury), would beknown or may be determined by a person of ordinary skill in the art. Theterm “treating” and conjugations thereof, include prevention of aninjury, pathology, condition, or disease (e.g. preventing thedevelopment of one or more symptoms of cancer (e.g. pancreatic cancer,breast cancer, multiple myeloma, cancers of secretory cells),neurodegenerative diseases, vanishing white matter disease, childhoodataxia with CNS hypo-myelination, and/or intellectual disabilitysyndromes (e.g. associated with impaired function of eIF2 or componentsin a signal transduction pathway including eIF2), or inflammatorydiseases (e.g. postsurgical cognitive dysfunction or traumatic braininjury),).

An “effective amount” is an amount sufficient to accomplish a statedpurpose (e.g. achieve the effect for which it is administered, treat adisease, reduce enzyme activity, increase enzyme activity, reduce one ormore symptoms of a disease or condition). An example of an “effectiveamount” is an amount sufficient to contribute to the treatment,prevention, or reduction of a symptom or symptoms of a disease, whichcould also be referred to as a “therapeutically effective amount.” A“reduction” of a symptom or symptoms (and grammatical equivalents ofthis phrase) means decreasing of the severity or frequency of thesymptom(s), or elimination of the symptom(s). A “prophylacticallyeffective amount” of a drug is an amount of a drug that, whenadministered to a subject, will have the intended prophylactic effect,e.g., preventing or delaying the onset (or reoccurrence) of an injury,disease, pathology or condition, or reducing the likelihood of the onset(or reoccurrence) of an injury, disease, pathology, or condition, ortheir symptoms. The full prophylactic effect does not necessarily occurby administration of one dose, and may occur only after administrationof a series of doses. Thus, a prophylactically effective amount may beadministered in one or more administrations. An “activity decreasingamount,” as used herein, refers to an amount of antagonist (inhibitor)required to decrease the activity of an enzyme or protein relative tothe absence of the antagonist. An “activity increasing amount,” as usedherein, refers to an amount of agonist (activator) required to increasethe activity of an enzyme or protein relative to the absence of theagonist. A “function disrupting amount,” as used herein, refers to theamount of antagonist (inhibitor) required to disrupt the function of anenzyme or protein relative to the absence of the antagonist. A “functionincreasing amount,” as used herein, refers to the amount of agonist(activator) required to increase the function of an enzyme or proteinrelative to the absence of the agonist. The exact amounts will depend onthe purpose of the treatment, and will be ascertainable by one skilledin the art using known techniques (see, e.g., Lieberman, PharmaceuticalDosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technologyof Pharmaceutical Compounding (1999); Pickar, Dosage Calculations(1999); and Remington: The Science and Practice of Pharmacy, 20thEdition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

The term “associated” or “associated with” in the context of a substanceor substance activity or function associated with a disease (e.g cancer(e.g. pancreatic cancer, breast cancer, multiple myeloma, or cancers ofsecretory cells), neurodegenerative diseases, vanishing white matterdisease, childhood ataxia with CNS hypo-myelination, and/or intellectualdisability syndromes (e.g. associated with impaired function of eIF2 orcomponents in a signal transduction pathway including eIF2)) means thatthe disease (e.g. cancer (e.g. pancreatic cancer, breast cancer,multiple myeloma, or cancers of secretory cells), neurodegenerativediseases, vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2)) is caused by (in whole or inpart), or a symptom of the disease is caused by (in whole or in part)the substance or substance activity or function. For example, a symptomof a disease or condition associated with an increase in eIF2α activitymay be a symptom that results (entirely or partially) from an increasein eIF2α activity (e.g increase in eIF2α phosphorylation or activity ofphosphorylated eIF2α or activity of eIF2α or increase in activity of aneIF2α signal transduction or signalling pathway). As used herein, whatis described as being associated with a disease, if a causative agent,could be a target for treatment of the disease. For example, a diseaseassociated with increased eIF2α activity or eIF2α pathway activity (e.g.phosphorylated eIF2α activity or pathway), may be treated with an agent(e.g. compound as described herein) effective for decreasing the levelof activity of eIF2α activity or eIF2α pathway or phosphorylated eIF2αactivity or pathway. For example, a disease associated withphosphorylated eIF2α, may be treated with an agent (e.g. compound asdescribed herein) effective for decreasing the level of activity ofphosphorylated eIF2α or a downstream component or effector ofphosphorylated eIF2α. For example, a disease associated with eIF2α, maybe treated with an agent (e.g. compound as described herein) effectivefor decreasing the level of activity of eIF2α or a downstream componentor effector of eIF2α.

“Control” or “control experiment” is used in accordance with its plainordinary meaning and refers to an experiment in which the subjects orreagents of the experiment are treated as in a parallel experimentexcept for omission of a procedure, reagent, or variable of theexperiment. In some instances, the control is used as a standard ofcomparison in evaluating experimental effects.

“Contacting” is used in accordance with its plain ordinary meaning andrefers to the process of allowing at least two distinct species (e.g.chemical compounds including biomolecules, or cells) to becomesufficiently proximal to react, interact or physically touch. It shouldbe appreciated, however, that the resulting reaction product can beproduced directly from a reaction between the added reagents or from anintermediate from one or more of the added reagents which can beproduced in the reaction mixture. The term “contacting” may includeallowing two species to react, interact, or physically touch, whereinthe two species may be a compound as described herein and a protein orenzyme (e.g. eIF2α or phosphorylated eIF2α or component of eIF2α pathwayor component of phosphorylated eIF2α pathway). In some embodimentscontacting includes allowing a compound described herein to interactwith a protein or enzyme that is involved in a signaling pathway (e.g.phosphorylated eIF2α pathway or eIF2α pathway).

As defined herein, the term “inhibition”, “inhibit”, “inhibiting” andthe like in reference to a protein-inhibitor (e.g. antagonist)interaction means negatively affecting (e.g. decreasing) the activity orfunction of the protein relative to the activity or function of theprotein in the absence of the inhibitor. In some embodiments inhibitionrefers to reduction of a disease or symptoms of disease. In someembodiments, inhibition refers to a reduction in the activity of asignal transduction pathway or signaling pathway. Thus, inhibitionincludes, at least in part, partially or totally blocking stimulation,decreasing, preventing, or delaying activation, or inactivating,desensitizing, or down-regulating signal transduction or enzymaticactivity or the amount of a protein. In some embodiments, inhibitionrefers to a decrease in the activity of a signal transduction pathway orsignaling pathway (e.g. eIF2α or phosphorylated eIF2α or eIF2α pathwayor phosphorylated eIF2α pathway or pathway activated by eIF2αphosphorylation). Thus, inhibition may include, at least in part,partially or totally decreasing stimulation, decreasing or reducingactivation, or inactivating, desensitizing, or down-regulating signaltransduction or enzymatic activity or the amount of a protein increasedin a disease (e.g. level of eIF2α activity or protein or level oractivity of a component of an eIF2α pathway or level of phosphorylatedeIF2α activity or protein or level or activity of a component of aphosphorylated eIF2α pathway, wherein each is associated with cancer(e.g. pancreatic cancer, breast cancer, multiple myeloma, or cancers ofsecretory cells), neurodegenerative diseases, vanishing white matterdisease, childhood ataxia with CNS hypo-myelination, and/or intellectualdisability syndromes (e.g. associated with impaired function of eIF2 orcomponents in a signal transduction pathway including eIF2)). Inhibitionmay include, at least in part, partially or totally decreasingstimulation, decreasing or reducing activation, or deactivating,desensitizing, or down-regulating signal transduction or enzymaticactivity or the amount of a protein (e.g. eIF2α, phosphorylated eIF2α,protein downstream in a pathway from eIF2α, protein downstream in apathway activated by phosphorylated eIF2α) that may modulate the levelof another protein or increase cell survival (e.g. decrease inphosphorylated eIF2α pathway activity may increase cell survival incells that may or may not have a increase in phosphorylated eIF2αpathway activity relative to a non-disease control or decrease in eIF2αpathway activity may increase cell survival in cells that may or may nothave a increase in eIF2α pathway activity relative to a non-diseasecontrol).

As defined herein, the term “activation”, “activate”, “activating” andthe like in reference to a protein-activator (e.g. agonist) interactionmeans positively affecting (e.g. increasing) the activity or function ofthe protein (e.g. eIF2α, phosphorylated eIF2α, component of pathwayincluding eIF2α, or component of pathway including phosphorylated eIF2α)relative to the activity or function of the protein in the absence ofthe activator (e.g. compound described herein). In some embodiments,activation refers to an increase in the activity of a signaltransduction pathway or signaling pathway (e.g. eIF2α or phosphorylatedeIF2α pathway). Thus, activation may include, at least in part,partially or totally increasing stimulation, increasing or enablingactivation, or activating, sensitizing, or up-regulating signaltransduction or enzymatic activity or the amount of a protein decreasedin a disease (e.g. level of eIF2α activity or level of protein oractivity decreased by phosphorylation of eIF2α or protein associatedwith cancer (e.g. pancreatic cancer, breast cancer, multiple myeloma, orcancers of secretory cells), neurodegenerative diseases, vanishing whitematter disease, childhood ataxia with CNS hypo-myelination, and/orintellectual disability syndromes (e.g. associated with impairedfunction of eIF2 or components in a signal transduction pathwayincluding eIF2)). Activation may include, at least in part, partially ortotally increasing stimulation, increasing or enabling activation, oractivating, sensitizing, or up-regulating signal transduction orenzymatic activity or the amount of a protein (e.g. eIF2α, proteindownstream of eIF2α, protein activated or upregulated by eIF2α, proteinactivated or upregulated by phosphorylation of eIF2α) that may modulatethe level of another protein or increase cell survival (e.g. increase ineIF2α activity may increase cell survival in cells that may or may nothave a reduction in eIF2α activity relative to a non-disease control).

The term “modulator” refers to a composition that increases or decreasesthe level of a target molecule or the function of a target molecule. Insome embodiments, a modulator of eIF2α or eIF2α pathway orphosphorylation of eIF2α or pathway activated by phorphorylation ofeIF2α is a compound that reduces the severity of one or more symptoms ofa disease associated with eIF2α or eIF2α pathway (e.g. diseaseassociated with an increase in the level of eIF2α activity or protein oreIF2α pathway activity or protein or eIF2α phorphorylation or pathwayactivated by eIF2α phosphorylation, for example cancer (e.g. pancreaticcancer, breast cancer, multiple myeloma, or cancers of secretory cells),neurodegenerative diseases, vanishing white matter disease, childhoodataxia with CNS hypo-myelination, and/or intellectual disabilitysyndromes (e.g. associated with impaired function of eIF2 or componentsin a signal transduction pathway including eIF2)) or a disease that isnot caused by eIF2α or eIF2α pathway but may benefit from modulation ofeIF2α or eIF2α pathway activity (e.g. decreasing in level or level ofactivity of eIF2α or eIF2α pathway). In embodiments, a modulator ofeIF2α or eIF2α pathway (e.g. phosphorylated eIF2α or phosphorylatedeIF2α pathway) is an anti-cancer agent. In embodiments, a modulator ofeIF2α or eIF2α pathway (e.g. phosphorylated eIF2α or phosphorylatedeIF2α pathway) is a neuroprotectant. In embodiments, a modulator ofeIF2α or eIF2α pathway (e.g. phosphorylated eIF2α or phosphorylatedeIF2α pathway) is a memory enhancing agent. In embodiments, a modulatorof eIF2α or eIF2α pathway is a long-term memory enhancing agent. Inembodiments, a modulator of eIF2α or eIF2α pathway (e.g. phosphorylatedeIF2α or phosphorylated eIF2α pathway) is a neuroprotective agent. Inembodiments, a modulator of eIF2α or eIF2α pathway (e.g. phosphorylatedeIF2α or phosphorylated eIF2α pathway) is an anti-inflammatory agent.

“Patient” or “subject in need thereof” refers to a living organismsuffering from or prone to a disease or condition that can be treated byadministration of a compound or pharmaceutical composition, as providedherein. Non-limiting examples include humans, other mammals, bovines,rats, mice, dogs, monkeys, goat, sheep, cows, deer, and othernon-mammalian animals. In some embodiments, a patient is human. In someembodiments, a patient is a domesticated animal. In some embodiments, apatient is a dog. In some embodiments, a patient is a parrot. In someembodiments, a patient is livestock animal. In some embodiments, apatient is a mammal. In some embodiments, a patient is a cat. In someembodiments, a patient is a horse. In some embodiments, a patient isbovine. In some embodiments, a patient is a canine. In some embodiments,a patient is a feline. In some embodiments, a patient is an ape. In someembodiments, a patient is a monkey. In some embodiments, a patient is amouse. In some embodiments, a patient is an experimental animal. In someembodiments, a patient is a rat. In some embodiments, a patient is ahamster. In some embodiments, a patient is a test animal. In someembodiments, a patient is a newborn animal. In some embodiments, apatient is a newborn human. In some embodiments, a patient is a newbornmammal. In some embodiments, a patient is an elderly animal. In someembodiments, a patient is an elderly human. In some embodiments, apatient is an elderly mammal. In some embodiments, a patient is ageriatric patient.

“Disease” or “condition” refer to a state of being or health status of apatient or subject capable of being treated with a compound,pharmaceutical composition, or method provided herein. In someembodiments, the disease is a disease related to (e.g. caused by) anincrease in the level of eIF2α, eIF2α phosphorylation, or eIF2α pathwayactivity, or pathway activated by phosphorylation of eIF2α. In someembodiments, the disease is a disease related to (e.g. caused by)neurodegeneration. In some embodiments, the disease is a disease relatedto (e.g. caused by) neural cell death. In some embodiments, the diseaseis a disease related to (e.g. caused by) a increase in the level ofeIF2α activity, eIF2α phosphorylation, eIF2α pathway activity, orphosphorylated eIF2α pathway activity. In some embodiments, the diseaseis cancer (e.g. pancreatic cancer, breast cancer, multiple myeloma, orcancers of secretory cells). In some embodiments, the disease is aneurodegenerative disease. In some embodiments, the disease is vanishingwhite matter disease. In some embodiments, the disease is childhoodataxia with CNS hypo-myelination. In some embodiments, the disease is anintellectual disability syndrome (e.g. associated with impaired functionof eIF2 or components in a signal transduction pathway including eIF2)).In some embodiments, the disease is an inflammatory disease (e.g.postoperative cognitive dysfunction or traumatic brain injury).

Examples of diseases, disorders, or conditions include, but are notlimited to, cancer (e.g. pancreatic cancer, breast cancer, multiplemyeloma, or cancers of secretory cells), neurodegenerative diseases,vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2). In some instances, “disease” or“condition” refers to cancer. In some further instances, “cancer” refersto human cancers and carcinomas, sarcomas, adenocarcinomas, lymphomas,leukemias, melanomas, etc., including solid and lymphoid cancers,kidney, breast, lung, bladder, colon, ovarian, prostate, pancreas,stomach, brain, head and neck, skin, uterine, testicular, glioma,esophagus, liver cancer, including hepatocarcinoma, lymphoma, includingB-acute lymphoblastic lymphoma, non-Hodgkin's lymphomas (e.g.,Burkitt's, Small Cell, and Large Cell lymphomas), Hodgkin's lymphoma,leukemia (including AML, ALL, and CML), and/or multiple myeloma. In somefurther instances, “cancer” refers to lung cancer, breast cancer,ovarian cancer, leukemia, lymphoma, melanoma, pancreatic cancer,sarcoma, bladder cancer, bone cancer, brain cancer, cervical cancer,colon cancer, esophageal cancer, gastric cancer, liver cancer, head andneck cancer, kidney cancer, myeloma, thyroid cancer, prostate cancer,metastatic cancer, or carcinoma. In embodiments, a disease isunsatisfactory long-term memory.

As used herein, the term “cancer” refers to all types of cancer,neoplasm or malignant tumors found in mammals, including leukemia,lymphoma, carcinomas and sarcomas. Exemplary cancers that may be treatedwith a compound, pharmaceutical composition, or method provided hereininclude lymphoma, sarcoma, bladder cancer, bone cancer, brain tumor,cervical cancer, colon cancer, esophageal cancer, gastric cancer, headand neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia,prostate cancer, breast cancer (e.g. ER positive, ER negative,chemotherapy resistant, herceptin resistant, HER2 positive, doxorubicinresistant, tamoxifen resistant, ductal carcinoma, lobular carcinoma,primary, metastatic), ovarian cancer, pancreatic cancer, liver cancer(e.g. hepatocellular carcinoma), lung cancer (e.g. non-small cell lungcarcinoma, squamous cell lung carcinoma, adenocarcinoma, large cell lungcarcinoma, small cell lung carcinoma, carcinoid, sarcoma), glioblastomamultiforme, glioma, or melanoma. Additional examples include, cancer ofthe thyroid, endocrine system, brain, breast, cervix, colon, head &neck, liver, kidney, lung, non-small cell lung, melanoma, mesothelioma,ovary, sarcoma, stomach, uterus or Medulloblastoma, Hodgkin's Disease,Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, glioma,glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primarythrombocytosis, primary macroglobulinemia, primary brain tumors, cancer,malignant pancreatic insulanoma, malignant carcinoid, urinary bladdercancer, premalignant skin lesions, testicular cancer, lymphomas, thyroidcancer, neuroblastoma, esophageal cancer, genitourinary tract cancer,malignant hypercalcemia, endometrial cancer, adrenal cortical cancer,neoplasms of the endocrine or exocrine pancreas, medullary thyroidcancer, medullary thyroid carcinoma, melanoma, colorectal cancer,papillary thyroid cancer, hepatocellular carcinoma, Paget's Disease ofthe Nipple, Phyllodes Tumors, Lobular Carcinoma, Ductal Carcinoma,cancer of the pancreatic stellate cells, cancer of the hepatic stellatecells, or prostate cancer.

The term “leukemia” refers broadly to progressive, malignant diseases ofthe blood-forming organs and is generally characterized by a distortedproliferation and development of leukocytes and their precursors in theblood and bone marrow. Leukemia is generally clinically classified onthe basis of (1) the duration and character of the disease-acute orchronic; (2) the type of cell involved; myeloid (myelogenous), lymphoid(lymphogenous), or monocytic; and (3) the increase or non-increase inthe number abnormal cells in the blood-leukemic or aleukemic(subleukemic). Exemplary leukemias that may be treated with a compound,pharmaceutical composition, or method provided herein include, forexample, acute nonlymphocytic leukemia, chronic lymphocytic leukemia,acute granulocytic leukemia, chronic granulocytic leukemia, acutepromyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, aleukocythemic leukemia, basophylic leukemia, blast cell leukemia, bovineleukemia, chronic myelocytic leukemia, leukemia cutis, embryonalleukemia, eosinophilic leukemia, Gross' leukemia, hairy-cell leukemia,hemoblastic leukemia, hemocytoblastic leukemia, histiocytic leukemia,stem cell leukemia, acute monocytic leukemia, leukopenic leukemia,lymphatic leukemia, lymphoblastic leukemia, lymphocytic leukemia,lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell leukemia,mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia,monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myeloidgranulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasmacell leukemia, multiple myeloma, plasmacytic leukemia, promyelocyticleukemia, Rieder cell leukemia, Schilling's leukemia, stem cellleukemia, subleukemic leukemia, or undifferentiated cell leukemia.

The term “sarcoma” generally refers to a tumor which is made up of asubstance like the embryonic connective tissue and is generally composedof closely packed cells embedded in a fibrillar or homogeneoussubstance. Sarcomas that may be treated with a compound, pharmaceuticalcomposition, or method provided herein include a chondrosarcoma,fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma, osteosarcoma,Abemethy's sarcoma, adipose sarcoma, liposarcoma, alveolar soft partsarcoma, ameloblastic sarcoma, botryoid sarcoma, chloroma sarcoma,chorio carcinoma, embryonal sarcoma, Wilms' tumor sarcoma, endometrialsarcoma, stromal sarcoma, Ewing's sarcoma, fascial sarcoma, fibroblasticsarcoma, giant cell sarcoma, granulocytic sarcoma, Hodgkin's sarcoma,idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcomaof B cells, lymphoma, immunoblastic sarcoma of T-cells, Jensen'ssarcoma, Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma,leukosarcoma, malignant mesenchymoma sarcoma, parosteal sarcoma,reticulocytic sarcoma, Rous sarcoma, serocystic sarcoma, synovialsarcoma, or telangiectaltic sarcoma.

The term “melanoma” is taken to mean a tumor arising from themelanocytic system of the skin and other organs. Melanomas that may betreated with a compound, pharmaceutical composition, or method providedherein include, for example, acral-lentiginous melanoma, amelanoticmelanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma,Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma,malignant melanoma, nodular melanoma, subungal melanoma, or superficialspreading melanoma.

The term “carcinoma” refers to a malignant new growth made up ofepithelial cells tending to infiltrate the surrounding tissues and giverise to metastases. Exemplary carcinomas that may be treated with acompound, pharmaceutical composition, or method provided herein include,for example, medullary thyroid carcinoma, familial medullary thyroidcarcinoma, acinar carcinoma, acinous carcinoma, adenocystic carcinoma,adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenalcortex, alveolar carcinoma, alveolar cell carcinoma, basal cellcarcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamouscell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma,bronchogenic carcinoma, cerebriform carcinoma, cholangiocellularcarcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma,corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinomacutaneum, cylindrical carcinoma, cylindrical cell carcinoma, ductcarcinoma, ductal carcinoma, carcinoma durum, embryonal carcinoma,encephaloid carcinoma, epiermoid carcinoma, carcinoma epithelialeadenoides, exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum,gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma,carcinoma gigantocellulare, glandular carcinoma, granulosa cellcarcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellularcarcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernephroidcarcinoma, infantile embryonal carcinoma, carcinoma in situ,intraepidermal carcinoma, intraepithelial carcinoma, Krompecher'scarcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma, lenticularcarcinoma, carcinoma lenticulare, lipomatous carcinoma, lobularcarcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullarycarcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma,carcinoma muciparum, carcinoma mucocellulare, mucoepidermoid carcinoma,carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes,nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans,osteoid carcinoma, papillary carcinoma, periportal carcinoma,preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma,renal cell carcinoma of kidney, reserve cell carcinoma, carcinomasarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinomascroti, signet-ring cell carcinoma, carcinoma simplex, small-cellcarcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cellcarcinoma, carcinoma spongiosum, squamous carcinoma, squamous cellcarcinoma, string carcinoma, carcinoma telangiectaticum, carcinomatelangiectodes, transitional cell carcinoma, carcinoma tuberosum,tubular carcinoma, tuberous carcinoma, verrucous carcinoma, or carcinomavillosum.

As used herein, the term “neurodegenerative disease” refers to a diseaseor condition in which the function of a subject's nervous system becomesimpaired. Examples of neurodegenerative diseases that may be treatedwith a compound, pharmaceutical composition, or method described hereininclude Alexander's disease, Alper's disease, Alzheimer's disease,Amyotrophic lateral sclerosis, Ataxia telangiectasia, Batten disease(also known as Spielmeyer-Vogt-Sjogren-Batten disease), Bovinespongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome,Corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporaldementia, Gerstmann-Sträussler-Scheinker syndrome, Huntington's disease,HIV-associated dementia, Kennedy's disease, Krabbe's disease, kuru, Lewybody dementia, Machado-Joseph disease (Spinocerebellar ataxia type 3),Multiple sclerosis, Multiple System Atrophy, Narcolepsy,Neuroborreliosis, Parkinson's disease, Pelizaeus-Merzbacher Disease,Pick's disease, Primary lateral sclerosis, Prion diseases, Refsum'sdisease, Sandhoff s disease, Schilder's disease, Subacute combineddegeneration of spinal cord secondary to Pernicious Anaemia,Schizophrenia, Spinocerebellar ataxia (multiple types with varyingcharacteristics), Spinal muscular atrophy, Steele-Richardson-Olszewskidisease, or Tabes dorsalis.

As used herein, the term “inflammatory disease” refers to a disease orcondition characterized by aberrant inflammation (e.g. an increasedlevel of inflammation compared to a control such as a healthy person notsuffering from a disease). Examples of inflammatory diseases includepostoperative cognitive dysfunction, traumatic brain injury, arthritis,rheumatoid arthritis, psoriatic arthritis, juvenile idiopathicarthritis, multiple sclerosis, systemic lupus erythematosus (SLE),myasthenia gravis, juvenile onset diabetes, diabetes mellitus type 1,Guillain-Barre syndrome, Hashimoto's encephalitis, Hashimoto'sthyroiditis, ankylosing spondylitis, psoriasis, Sjogren's syndrome,vasculitis, glomerulonephritis, auto-immune thyroiditis, Behcet'sdisease, Crohn's disease, ulcerative colitis, bullous pemphigoid,sarcoidosis, ichthyosis, Graves ophthalmopathy, inflammatory boweldisease, Addison's disease, Vitiligo, asthma, allergic asthma, acnevulgaris, celiac disease, chronic prostatitis, inflammatory boweldisease, pelvic inflammatory disease, reperfusion injury, sarcoidosis,transplant rejection, interstitial cystitis, atherosclerosis, and atopicdermatitis. Proteins associated with inflammation and inflammatorydiseases (e.g. aberrant expression being a symptom or cause or marker ofthe disease) include interleukin-6 (IL-6), interleukin-8 (IL-8),interleukin-18 (IL-18), TNF-α (tumor necrosis factor-alpha), andC-reactive protein (CRP).

The term “postoperative cognitive dysfunction” refers to a decline incognitive function (e.g. memory or executive function (e.g. workingmemory, reasoning, task flexibility, speed of processing, or problemsolving)) following surgery.

The term “signaling pathway” as used herein refers to a series ofinteractions between cellular and optionally extra-cellular components(e.g. proteins, nucleic acids, small molecules, ions, lipids) thatconveys a change in one component to one or more other components, whichin turn may convey a change to additional components, which isoptionally propagated to other signaling pathway components.

“Pharmaceutically acceptable excipient” and “pharmaceutically acceptablecarrier” refer to a substance that aids the administration of an activeagent to and absorption by a subject and can be included in thecompositions of the present invention without causing a significantadverse toxicological effect on the patient. Non-limiting examples ofpharmaceutically acceptable excipients include water, NaCl, normalsaline solutions, lactated Ringer's, normal sucrose, normal glucose,binders, fillers, disintegrants, lubricants, coatings, sweeteners,flavors, salt solutions (such as Ringer's solution), alcohols, oils,gelatins, carbohydrates such as lactose, amylose or starch, fatty acidesters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, andthe like. Such preparations can be sterilized and, if desired, mixedwith auxiliary agents such as lubricants, preservatives, stabilizers,wetting agents, emulsifiers, salts for influencing osmotic pressure,buffers, coloring, and/or aromatic substances and the like that do notdeleteriously react with the compounds of the invention. One of skill inthe art will recognize that other pharmaceutical excipients are usefulin the present invention.

The term “preparation” is intended to include the formulation of theactive compound with encapsulating material as a carrier providing acapsule in which the active component with or without other carriers, issurrounded by a carrier, which is thus in association with it.Similarly, cachets and lozenges are included. Tablets, powders,capsules, pills, cachets, and lozenges can be used as solid dosage formssuitable for oral administration.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact, intravenous,parenteral, intraperitoneal, intramuscular, intralesional, intrathecal,intracranial, intranasal or subcutaneous administration, or theimplantation of a slow-release device, e.g., a mini-osmotic pump, to asubject. Administration is by any route, including parenteral andtransmucosal (e.g., buccal, sublingual, palatal, gingival, nasal,vaginal, rectal, or transdermal). Parenteral administration includes,e.g., intravenous, intramuscular, intra-arteriole, intradermal,subcutaneous, intraperitoneal, intraventricular, and intracranial. Othermodes of delivery include, but are not limited to, the use of liposomalformulations, intravenous infusion, transdermal patches, etc. By“co-administer” it is meant that a composition described herein isadministered at the same time, just prior to, or just after theadministration of one or more additional therapies (e.g. anti-canceragent, chemotherapeutic, or treatment for a neurodegenerative disease).The compound of the invention can be administered alone or can becoadministered to the patient. Coadministration is meant to includesimultaneous or sequential administration of the compound individuallyor in combination (more than one compound or agent). Thus, thepreparations can also be combined, when desired, with other activesubstances (e.g. to reduce metabolic degradation). The compositions ofthe present invention can be delivered by transdermally, by a topicalroute, formulated as applicator sticks, solutions, suspensions,emulsions, gels, creams, ointments, pastes, jellies, paints, powders,and aerosols. Oral preparations include tablets, pills, powder, dragees,capsules, liquids, lozenges, cachets, gels, syrups, slurries,suspensions, etc., suitable for ingestion by the patient. Solid formpreparations include powders, tablets, pills, capsules, cachets,suppositories, and dispersible granules. Liquid form preparationsinclude solutions, suspensions, and emulsions, for example, water orwater/propylene glycol solutions. The compositions of the presentinvention may additionally include components to provide sustainedrelease and/or comfort. Such components include high molecular weight,anionic mucomimetic polymers, gelling polysaccharides and finely-divideddrug carrier substrates. These components are discussed in greaterdetail in U.S. Pat. Nos. 4,911,920; 5,403,841; 5,212,162; and 4,861,760.The entire contents of these patents are incorporated herein byreference in their entirety for all purposes. The compositions of thepresent invention can also be delivered as microspheres for slow releasein the body. For example, microspheres can be administered viaintradermal injection of drug-containing microspheres, which slowlyrelease subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645,1995; as biodegradable and injectable gel formulations (see, e.g., GaoPharm. Res. 12:857-863, 1995); or, as microspheres for oraladministration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674,1997). In another embodiment, the formulations of the compositions ofthe present invention can be delivered by the use of liposomes whichfuse with the cellular membrane or are endocytosed, i.e., by employingreceptor ligands attached to the liposome, that bind to surface membraneprotein receptors of the cell resulting in endocytosis. By usingliposomes, particularly where the liposome surface carries receptorligands specific for target cells, or are otherwise preferentiallydirected to a specific organ, one can focus the delivery of thecompositions of the present invention into the target cells in vivo.(See, e.g., Al-Muhammed, J. Microencapsul. 13:293-306, 1996; Chonn,Curr. Opin. Biotechnol. 6:698-708, 1995; Ostro, Am. J. Hosp. Pharm.46:1576-1587, 1989). The compositions of the present invention can alsobe delivered as nanoparticles.

Pharmaceutical compositions provided by the present invention includecompositions wherein the active ingredient (e.g. compounds describedherein, including embodiments or examples) is contained in atherapeutically effective amount, i.e., in an amount effective toachieve its intended purpose. The actual amount effective for aparticular application will depend, inter alia, on the condition beingtreated. When administered in methods to treat a disease, suchcompositions will contain an amount of active ingredient effective toachieve the desired result, e.g., modulating the activity of a targetmolecule (e.g. eIF2α or component of eIF2α signal transduction pathwayor component of phosphorylated eIF2α pathway), and/or reducing,eliminating, or slowing the progression of disease symptoms (e.g.symptoms of cancer (e.g. pancreatic cancer, breast cancer, multiplemyeloma, or cancers of secretory cells), neurodegenerative diseases,vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2)). Determination of atherapeutically effective amount of a compound of the invention is wellwithin the capabilities of those skilled in the art, especially in lightof the detailed disclosure herein.

The dosage and frequency (single or multiple doses) administered to amammal can vary depending upon a variety of factors, for example,whether the mammal suffers from another disease, and its route ofadministration; size, age, sex, health, body weight, body mass index,and diet of the recipient; nature and extent of symptoms of the diseasebeing treated (e.g. symptoms of cancer (e.g. pancreatic cancer, breastcancer, multiple myeloma, or cancers of secretory cells),neurodegenerative diseases, vanishing white matter disease, childhoodataxia with CNS hypo-myelination, and/or intellectual disabilitysyndromes (e.g. associated with impaired function of eIF2 or componentsin a signal transduction pathway including eIF2)), kind of concurrenttreatment, complications from the disease being treated or otherhealth-related problems. Other therapeutic regimens or agents can beused in conjunction with the methods and compounds of Applicants'invention. Adjustment and manipulation of established dosages (e.g.,frequency and duration) are well within the ability of those skilled inthe art.

For any compound described herein, the therapeutically effective amountcan be initially determined from cell culture assays. Targetconcentrations will be those concentrations of active compound(s) thatare capable of achieving the methods described herein, as measured usingthe methods described herein or known in the art.

As is well known in the art, therapeutically effective amounts for usein humans can also be determined from animal models. For example, a dosefor humans can be formulated to achieve a concentration that has beenfound to be effective in animals. The dosage in humans can be adjustedby monitoring compounds effectiveness and adjusting the dosage upwardsor downwards, as described above. Adjusting the dose to achieve maximalefficacy in humans based on the methods described above and othermethods is well within the capabilities of the ordinarily skilledartisan.

Dosages may be varied depending upon the requirements of the patient andthe compound being employed. The dose administered to a patient, in thecontext of the present invention should be sufficient to effect abeneficial therapeutic response in the patient over time. The size ofthe dose also will be determined by the existence, nature, and extent ofany adverse side-effects. Determination of the proper dosage for aparticular situation is within the skill of the practitioner. Generally,treatment is initiated with smaller dosages which are less than theoptimum dose of the compound. Thereafter, the dosage is increased bysmall increments until the optimum effect under circumstances isreached.

Dosage amounts and intervals can be adjusted individually to providelevels of the administered compound effective for the particularclinical indication being treated. This will provide a therapeuticregimen that is commensurate with the severity of the individual'sdisease state.

Utilizing the teachings provided herein, an effective prophylactic ortherapeutic treatment regimen can be planned that does not causesubstantial toxicity and yet is effective to treat the clinical symptomsdemonstrated by the particular patient. This planning should involve thecareful choice of active compound by considering factors such ascompound potency, relative bioavailability, patient body weight,presence and severity of adverse side effects, preferred mode ofadministration and the toxicity profile of the selected agent.

The compounds described herein can be used in combination with oneanother, with other active agents known to be useful in treating cancer(e.g. pancreatic cancer, breast cancer, multiple myeloma, or cancers ofsecretory cells), neurodegenerative diseases, vanishing white matterdisease, childhood ataxia with CNS hypo-myelination, and/or intellectualdisability syndromes (e.g. associated with impaired function of eIF2 orcomponents in a signal transduction pathway including eIF2), or withadjunctive agents that may not be effective alone, but may contribute tothe efficacy of the active agent.

In some embodiments, co-administration includes administering one activeagent within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of a secondactive agent. Co-administration includes administering two active agentssimultaneously, approximately simultaneously (e.g., within about 1, 5,10, 15, 20, or 30 minutes of each other), or sequentially in any order.In some embodiments, co-administration can be accomplished byco-formulation, i.e., preparing a single pharmaceutical compositionincluding both active agents. In other embodiments, the active agentscan be formulated separately. In another embodiment, the active and/oradjunctive agents may be linked or conjugated to one another. In someembodiments, the compounds described herein may be combined withtreatments for cancer (e.g. pancreatic cancer, breast cancer, multiplemyeloma, or cancers of secretory cells), neurodegenerative diseases,vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2), or inflammatory diseases (e.g.POCD or TBI), such as surgery.

The term “eIF2alpha” or “eIF2α” refers to the protein “Eukaryotictranslation initiation factor 2A”. In embodiments, “eIF2alpha” or“eIF2α” refers to the human protein. Included in the term “eIF2alpha” or“eIF2α” are the wildtype and mutant forms of the protein. Inembodiments, “eIF2alpha” or “eIF2α” refers to the protein associatedwith Entrez Gene 83939, OMIM 609234, UniProt Q9BY44, and/or RefSeq(protein) NP_114414. In embodiments, the reference numbers immediatelyabove refer to the protein, and associated nucleic acids, known as ofthe date of filing of this application.

“Anti-cancer agent” is used in accordance with its plain ordinarymeaning and refers to a composition (e.g. compound, drug, antagonist,inhibitor, modulator) having antineoplastic properties or the ability toinhibit the growth or proliferation of cells. In some embodiments, ananti-cancer agent is a chemotherapeutic. In some embodiments, ananti-cancer agent is an agent identified herein having utility inmethods of treating cancer. In some embodiments, an anti-cancer agent isan agent approved by the FDA or similar regulatory agency of a countryother than the USA, for treating cancer. Examples of anti-cancer agentsinclude, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2)inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib/AZD6244,GSK1120212/trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901,U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylatingagents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan,melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogenmustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil,meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine,thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine, lomusitne, semustine, streptozocin), triazenes(decarbazine)), anti-metabolites (e.g., 5-azathioprine, leucovorin,capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folicacid analog (e.g., methotrexate), or pyrimidine analogs (e.g.,fluorouracil, floxouridine, Cytarabine), purine analogs (e.g.,mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g.,vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin,paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g.,irinotecan, topotecan, amsacrine, etoposide (VP16), etoposide phosphate,teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin,daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin,mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g.cisplatin, oxaloplatin, carboplatin), anthracenedione (e.g.,mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazinederivative (e.g., procarbazine), adrenocortical suppressant (e.g.,mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide),antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g.,L-asparaginase), inhibitors of mitogen-activated protein kinasesignaling (e.g. U0126, PD98059, PD184352, PD0325901, ARRY-142886,SB239063, SP600125, BAY 43-9006, wortmannin, or LY294002, Sykinhibitors, mTOR inhibitors, antibodies (e.g., rituxan), gossyphol,genasense, polyphenol E, Chlorofusin, all trans-retinoic acid (ATRA),bryostatin, tumor necrosis factor-related apoptosis-inducing ligand(TRAIL), 5-aza-2′-deoxycytidine, all trans retinoic acid, doxorubicin,vincristine, etoposide, gemcitabine, imatinib (GLEEVEC®), geldanamycin,17-N-Allylamino-17-Demethoxygeldanamycin (17-AAG), flavopiridol,LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, PD184352,20-epi-1, 25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone;aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TKantagonists; altretamine; ambamustine; amidox; amifostine;aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;andrographolide; angiogenesis inhibitors; antagonist D; antagonist G;antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen,prostatic carcinoma; antiestrogen; antineoplaston; antisenseoligonucleotides; aphidicolin glycinate; apoptosis gene modulators;apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; argininedeaminase; asulacrine; atamestane; atrimustine; axinastatin 1;axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatinIII derivatives; balanol; batimastat; BCR/ABL antagonists;benzochlorins; benzoylstaurosporine; beta lactam derivatives;beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistrateneA; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2;capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol;dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA;ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene;emitefur; epirubicin; epristeride; estramustine analogue; estrogenagonists; estrogen antagonists; etanidazole; etoposide phosphate;exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride;flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin;pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine;pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin;pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenyl acetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen-binding protein; sizofuran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatinstimalamer, Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin,acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate;aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride;decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate;diaziquone; doxorubicin; doxorubicin hydrochloride; droloxifene;droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate;eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate;epipropidine; epirubicin hydrochloride; erbulozole; esorubicinhydrochloride; estramustine; estramustine phosphate sodium; etanidazole;etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride;fazarabine; fenretinide; floxuridine; fludarabine phosphate;fluorouracil; fluorocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; iimofosine; interleukin (includingrecombinant interleukin II, or rlL.sub.2), interferon alfa-2a;interferon alfa-2b; interferon alfa-nl; interferon alfa-n3; interferonbeta-1a; interferon gamma-1b; iproplatin; irinotecan hydrochloride;lanreotide acetate; letrozole; leuprolide acetate; liarozolehydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride;masoprocol; maytansine; mechlorethamine hydrochloride; megestrolacetate; melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie;nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol;safingol hydrochloride; semustine; simtrazene; sparfosate sodium;sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride, agents that arrest cells in the G2-M phases and/ormodulate the formation or stability of microtubules, (e.g. TAXOL™ (i.e.paclitaxel), TAXOTERE™, compounds comprising the taxane skeleton,Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128),Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829,Discodermolide (i.e. as NVP-XX-A-296), ABT-751 (Abbott, i.e. E-7010),Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g.Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4,Spongistatin 5, Spongistatin 6, Spongistatin 7, Spongistatin 8, andSpongistatin 9), Cemadotin hydrochloride (i.e. LU-103793 andNSC-D-669356), Epothilones (e.g. Epothilone A, Epothilone B, EpothiloneC (i.e. desoxyepothilone A or dEpoA), Epothilone D (i.e. KOS-862, dEpoB,and desoxyepothilone B), Epothilone E, Epothilone F, Epothilone BN-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21-aminoepothilone B(i.e. BMS-310705), 21-hydroxyepothilone D (i.e. Desoxyepothilone F anddEpoF), 26-fluoroepothilone, Auristatin PE (i.e. NSC-654663), Soblidotin(i.e. TZT-1027), LS-4559-P (Pharmacia, i.e. LS-4577), LS-4578(Pharmacia, i.e. LS-477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia),RPR-112378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877(Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2(Hungarian Academy of Sciences), BSF-223651 (BASF, i.e. ILX-651 andLU-223651), SAH-49960 (Lilly/Novartis), SDZ-268970 (Lilly/Novartis),AM-97 (Armad/Kyowa Hakko), AM-132 (Armad), AM-138 (Armad/Kyowa Hakko),IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto,i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e. AVE-8062,AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A), Vitilevuamide, TubulysinA, Canadensol, Centaureidin (i.e. NSC-106969), T-138067 (Tularik, i.e.T-67, TL-138067 and TI-138067), COBRA-1 (Parker Hughes Institute, i.e.DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas StateUniversity), Oncocidin Al (i.e. BTO-956 and DIME), DDE-313 (ParkerHughes Institute), Fijianolide B, Laulimalide, SPA-2 (Parker HughesInstitute), SPA-1 (Parker Hughes Institute, i.e. SPIKET-P), 3-IAABU(Cytoskeleton/Mt. Sinai School of Medicine, i.e. MF-569), Narcosine(also known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972(Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton/Mt. Sinai School ofMedicine, i.e. MF-191), TMPN (Arizona State University), Vanadoceneacetylacetonate, T-138026 (Tularik), Monsatrol, lnanocine (i.e.NSC-698666), 3-IAABE (Cytoskeleton/Mt. Sinai School of Medicine),A-204197 (Abbott), T-607 (Tuiarik, i.e. T-900607), RPR-115781 (Aventis),Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin,lsoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin,Halichondrin B, D-64131 (Asta Medica), D-68144 (Asta Medica),Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A,TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, (−)-Phenylahistin(i.e. NSCL-96F037), D-68838 (Asta Medica), D-68836 (Asta Medica),Myoseverin B, D-43411 (Zentaris, i.e. D-81862), A-289099 (Abbott),A-318315 (Abbott), HTI-286 (i.e. SPA-110, trifluoroacetate salt)(Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC-12983 (NCI),Resverastatin phosphate sodium, BPR-OY-007 (National Health ResearchInstitutes), and SSR-250411 (Sanofi)), steroids (e.g., dexamethasone),finasteride, aromatase inhibitors, gonadotropin-releasing hormoneagonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids(e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate,megestrol acetate, medroxyprogesterone acetate), estrogens (e.g.,diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen),androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogen(e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guérin(BCG), levamisole, interleukin-2, alpha-interferon, etc.), monoclonalantibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, andanti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33monoclonal antibody-calicheamicin conjugate, anti-CD22 monoclonalantibody-pseudomonas exotoxin conjugate, etc.), radioimmunotherapy(e.g., anti-CD20 monoclonal antibody conjugated to ¹¹¹In, ⁹⁰Y, or ¹³¹I,etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin,epirubicin, topotecan, itraconazole, vindesine, cerivastatin,vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan,clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib,gefitinib, EGFR inhibitors, epidermal growth factor receptor(EGFR)-targeted therapy or therapeutic (e.g. gefitinib (Iressa™)erlotinib (Tarceva™), cetuximab (Erbitux™), lapatinib (Tykerb™),panitumumab (Vectibix™) vandetanib (Caprelsa™), afatinib/BIBW2992,CI-1033/canertinib, neratinib/HKI-272, CP-724714, TAK-285, AST-1306,ARRY334543, ARRY-380, AG-1478, dacomitinib/PF299804, OSI-420/desmethylerlotinib, AZD8931, AEE788, pelitinib/EKB-569, CUDC-101, WZ8040, WZ4002,WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib,sunitinib, dasatinib, or the like.

“Chemotherapeutic” or “chemotherapeutic agent” is used in accordancewith its plain ordinary meaning and refers to a chemical composition orcompound having antineoplastic properties or the ability to inhibit thegrowth or proliferation of cells.

Additionally, the compounds described herein can be co-administered withconventional immunotherapeutic agents including, but not limited to,immunostimulants (e.g., Bacillus Calmette-Guérin (BCG), levamisole,interleukin-2, alpha-interferon, etc.), monoclonal antibodies (e.g.,anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonalantibodies), immunotoxins (e.g., anti-CD33 monoclonalantibody-calicheamicin conjugate, anti-CD22 monoclonalantibody-pseudomonas exotoxin conjugate, etc.), and radioimmunotherapy(e.g., anti-CD20 monoclonal antibody conjugated to ¹¹¹In, ⁹⁰Y or ¹³¹I,etc.).

In a further embodiment, the compounds described herein can beco-administered with conventional radiotherapeutic agents including, butnot limited to, radionuclides such as ⁴⁷Sc, ⁶⁴Cu, ⁶⁷Cu, ⁸⁹Sr, ⁸⁶Y, ⁸⁷Y,⁹⁰Y, ¹⁰⁵Rh, ¹¹¹Ag, ¹¹¹In, ^(117m)sn, ¹⁴⁹Pm, ¹⁵³Sm, ¹⁶⁶Ho, ¹⁷⁷Lu, ¹⁸⁶Re,¹⁸⁸Re, ²¹¹At, and ²¹²Bi, optionally conjugated to antibodies directedagainst tumor antigens.

Methods of Treatment

In a first aspect is provided a method of treating an integrated stressresponse-associated disease in a patient in need of such treatment, themethod including administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, to the patient,wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; le, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In embodiments, the integrated stress response-associated disease iscancer. In embodiments, the integrated stress response-associateddisease is a neurodegenerative disease. In embodiments, the integratedstress response-associated disease is vanishing white matter disease. Inembodiments, the integrated stress response-associated disease ischildhood ataxia with CNS hypo-myelination. In embodiments, theintegrated stress response-associated disease is an intellectualdisability syndrome.

In another aspect is provided a method of treating a disease associatedwith phosphorylation of eIF2α in a patient in need of such treatment,the method including administering a therapeutically effective amount ofa compound, or a pharmaceutically acceptable salt thereof, to thepatient, wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In embodiments, the disease associated with phosphorylation of eIF2α iscancer. In embodiments, the disease associated with phosphorylation ofeIF2α is a neurodegenerative disease. In embodiments, the diseaseassociated with phosphorylation of eIF2α is vanishing white matterdisease. In embodiments, the disease associated with phosphorylation ofeIF2α is childhood ataxia with CNS hypo-myelination. In embodiments, thedisease associated with phosphorylation of eIF2α is an intellectualdisability syndrome

In another aspect is provided a method of treating a disease in apatient in need of such treatment, the method including administering atherapeutically effective amount of a compound to the patient, whereinthe disease is selected from the group consisting of cancer, aneurodegenerative disease, vanishing white matter disease, childhoodataxia with CNS hypo-myelination, and an intellectual disabilitysyndrome; and wherein the compound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In embodiments, the disease is cancer. In embodiments, the disease is aneurodegenerative disease. In embodiments, the disease is vanishingwhite matter disease. In embodiments, the disease is childhood ataxiawith CNS hypo-myelination. In embodiments, the disease is anintellectual disability syndrome

In another aspect is provided a method of treating an inflammatorydisease in a patient in need of such treatment, the method includingadministering a therapeutically effective amount of a compound, or apharmaceutically acceptable salt thereof, to the patient, wherein thecompound has the formula:

Ring A is substituted or unsubstituted cycloalkylene or substituted orunsubstituted arylene. L¹, L², L³, and L⁴ are independently a bond,—NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkyleneor substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

In embodiments, the inflammatory disease is associated with neurologicalinflammation. In embodiments, the inflammatory disease is postoperativecognitive dysfunction. In embodiments, the inflammatory disease istraumatic brain injury.

The embodiments described herein below may be applied to any of themethods of treatment described herein.

In embodiments, ring A is substituted or unsubstituted cycloalkylene. Inembodiments, ring A is substituted or unsubstituted C₃-C₈ cycloalkylene.In embodiments, ring A is substituted or unsubstituted C₃-C₆cycloalkylene. In embodiments, ring A is substituted or unsubstitutedC₃-C₄ cycloalkylene. In embodiments, ring A is substituted orunsubstituted C₄-C₈ cycloalkylene. In embodiments, ring A is substitutedor unsubstituted C₄-C₆ cycloalkylene. In embodiments, ring A issubstituted or unsubstituted cyclohexylene. In embodiments, ring A issubstituted or unsubstituted cyclobutylene. In embodiments, ring A issubstituted or unsubstituted cyclopentylene. In embodiments, ring A issubstituted or unsubstituted C₄-C₆ cycloalkenylene. In embodiments, ringA is unsubstituted cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₈ cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₆ cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₄ cycloalkylene. In embodiments, ring A isunsubstituted C₄-C₈ cycloalkylene. In embodiments, ring A isunsubstituted C₄-C₆ cycloalkylene. In embodiments, ring A isunsubstituted cyclohexylene. In embodiments, ring A is unsubstitutedcyclobutylene. In embodiments, ring A is unsubstituted cyclopentylene.In embodiments, ring A is unsubstituted C₄-C₆ cycloalkenylene. Inembodiments, ring A is substituted or unsubstituted arylene. Inembodiments, ring A is substituted or unsubstituted C₆-C₁₀ arylene. Inembodiments, ring A is substituted or unsubstituted phenylene. Inembodiments, ring A is substituted or unsubstituted naphthylene. Inembodiments, ring A is unsubstituted C₆-C₁₀ arylene. In embodiments,ring A is unsubstituted phenylene. In embodiments, ring A isunsubstituted naphthylene. It is understood that when ring A isunsubstituted, it does not include additional substituents in additionto the bonds explicitly shown in the formula of interest (e.g. formulaI, Ia, etc.).

L¹ may be a bond or substituted or unsubstituted alkylene. L¹ may besubstituted or unsubstituted C₁-C₅ alkylene. L¹ may be substituted orunsubstituted C₁-C₃ alkylene. L¹ may be substituted or unsubstitutedmethylene. L¹ may be a bond. L¹ may be an unsubstituted alkylene. L¹ maybe an unsubstituted methylene. L¹ may be an unsubstituted ethylene. L¹may be a methylene substituted with an unsubstituted alkyl L¹ may be amethylene substituted with an unsubstituted C₁-C₄ alkyl L¹ may be amethylene substituted with an unsubstituted C₁-C₃ alkyl.

L³ may be a bond or substituted or unsubstituted alkylene. L³ may besubstituted or unsubstituted C₁-C₅ alkylene. L³ may be substituted orunsubstituted C₁-C₃ alkylene. L³ may be substituted or unsubstitutedmethylene. L³ may be a bond. L³ may be an unsubstituted alkylene. L³ maybe an unsubstituted methylene. L³ may be an unsubstituted ethylene. L³may be a methylene substituted with an unsubstituted alkyl L³ may be amethylene substituted with an unsubstituted C₁-C₄ alkyl L³ may be amethylene substituted with an unsubstituted C₁-C₃ alkyl. In embodiments,L¹ and L³ may be a bond. In embodiments, L¹ and L³ may independently bean unsubstituted alkylene. In embodiments, L¹ and L³ may be anunsubstituted methylene.

In embodiments, R¹ is hydrogen. In embodiments, R¹ is —CH₂CCH. Inembodiments, R¹ is

In embodiments, R¹ is

In embodiments, R¹ is

In embodiments, R¹ is substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. Inembodiments, R¹ is substituted or unsubstituted alkyl. In embodiments,R¹ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R¹ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R¹ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R¹ isunsubstituted alkyl. In embodiments, R¹ is unsubstituted C₁-C₈ alkyl. Inembodiments, R¹ is unsubstituted C₁-C₆ alkyl. In embodiments, le isunsubstituted C₁-C₄ alkyl. In embodiments, R¹ is substituted orunsubstituted heteroalkyl. In embodiments, R¹ is substituted orunsubstituted 2 to 8 membered heteroalkyl. In embodiments, le isunsubstituted 2 to 8 membered heteroalkyl.

In embodiments, R³ is hydrogen. In embodiments, R³ is —CH₂CCH. Inembodiments, R³ is

In embodiments, R³ is

In embodiments, R³ is

In embodiments, R³ is substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. Inembodiments, R³ is substituted or unsubstituted alkyl. In embodiments,R³ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R³ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R³ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R³ isunsubstituted alkyl. In embodiments, R³ is unsubstituted C₁-C₈ alkyl. Inembodiments, R³ is unsubstituted C₁-C₆ alkyl. In embodiments, R³ isunsubstituted C₁-C₄ alkyl. In embodiments, R³ is substituted orunsubstituted heteroalkyl. In embodiments, R³ is substituted orunsubstituted 2 to 8 membered heteroalkyl. In embodiments, R³ isunsubstituted 2 to 8 membered heteroalkyl.

In embodiments, R⁵ is independently

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO ₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R⁵ isindependentlyhalogen, —OCH₃, —OCH₂Ph, —CH₃, —OH, —CF₃, —CCl₃, —CN, —S(O)CH₃, —NO₂,—C(O)CH₃, —C(O)Ph, —CH(CH₃)₂, —CCSi(CH₃)₃, or —CCH. In embodiments, R⁵is —F. In embodiments, R⁵ is —Cl. In embodiments, R⁵ is —Br. Inembodiments, R⁵ is —I. In embodiments, R⁵ is substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. In embodiments, R⁵ is unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. Inembodiments, R⁵ is —OCH₃. In embodiments, R⁵ is —OCH₂Ph. In embodiments,R⁵ is —CH₃. In embodiments, R⁵ is —OH. In embodiments, R⁵ is —CF₃. Inembodiments, R⁵ is —CCl₃. In embodiments, R⁵ is —CN. In embodiments, R⁵is —S(O)CH₃. In embodiments, R⁵ is —NO₂. In embodiments, R⁵ is —C(O)CH₃.In embodiments, R⁵ is —C(O)Ph. In embodiments, R⁵ is —CH(CH₃)₂. Inembodiments, R⁵ is —CCSi(CH₃)₃. In embodiments, R⁵ is —C(NN)CF₃. Inembodiments, R⁵ is —C(NH—NH)CF₃. In embodiments, R⁵ is —CCH. Inembodiments, R⁵ is —CH₂CCH. In embodiments, R⁵ is —SH. In embodiments,R⁵ is —SO₂Cl. In embodiments, R⁵ is —SO₃H. In embodiments, R⁵ is —SO₄H.In embodiments, R⁵ is —SO₂NH₂. In embodiments, R⁵ is —NHNH₂. Inembodiments, R⁵ is —ONH₂. In embodiments, R⁵ is —NHC═(O)NHNH₂. Inembodiments, R⁵ is —NHC═(O)NH₂. In embodiments, R⁵ is —NHSO₂H. Inembodiments, R⁵ is —NHC═(O)H. In embodiments, R⁵ is —NHC(O)OH. Inembodiments, R⁵ is —NHOH. In embodiments, R⁵ is —OCH₃. In embodiments,R⁵ is —OCF₃. In embodiments, R⁵ is —OCHF₂. In embodiments, R⁵ is —N₃. Inembodiments, R⁵ is

In embodiments, R⁶ is independently halogen, —OCH₃, —OCH₂Ph, —C(O)Ph,—CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂,—C(O)CH₃, —CH(CH₃)₂, —CCS i(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H,—SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCHF₂,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R⁶ isindependentlyhalogen, —OCH₃, —OCH₂Ph, —CH₃, —OH, —CF₃, —CCl₃, —CN, —S(O)CH₃, —NO₂,—C(O)CH₃, —C(O)Ph, —CH(CH₃)₂, —CCSi(CH₃)₃, or —CCH. In embodiments, R⁶is —F. In embodiments, R⁶ is —Cl. In embodiments, R⁶ is —Br. Inembodiments, R⁶ is —I. In embodiments, R⁶ is substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. In embodiments, R⁶ is unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. Inembodiments, R⁶ is —OCH₃. In embodiments, R⁶ is —OCH₂Ph. In embodiments,R⁶ is —CH₃. In embodiments, R⁶ is —OH. In embodiments, R⁶ is —CF₃. Inembodiments, R⁶ is —CCl₃. In embodiments, R⁶ is —CN. In embodiments, R⁶is —S(O)CH₃. In embodiments, R⁶ is —NO₂. In embodiments, R⁶ is —C(O)CH₃.In embodiments, R⁶ is —C(O)Ph. In embodiments, R⁶ is —CH(CH₃)₂. Inembodiments, R⁶ is —CCSi(CH₃)₃. In embodiments, R⁶ is —C(NN)CF₃. Inembodiments, R⁶ is —C(NH—NH)CF₃. In embodiments, R⁶ is —CCH. Inembodiments, R⁶ is —CH₂CCH. In embodiments, R⁶ is —SH. In embodiments,R⁶ is —SO₂Cl. In embodiments, R⁶ is —SO₃H. In embodiments, R⁶ is —SO₄H.In embodiments, R⁶ is —SO₂NH₂. In embodiments, R⁶ is —NHNH₂. Inembodiments, R⁶ is —ONH₂. In embodiments, R⁶ is —NHC═(O)NHNH₂. Inembodiments, R⁶ is —NHC═(O)NH₂. In embodiments, R⁶ is —NHSO₂H. Inembodiments, R⁶ is —NHC═(O)H. In embodiments, R⁶ is —NHC(O)OH. Inembodiments, R⁶ is —NHOH. In embodiments, R⁶ is —OCH₃. In embodiments,R⁶ is —OCF₃. In embodiments, R⁶ is —OCHF₂. In embodiments, R⁶ is —N₃. Inembodiments, R⁶ is

In embodiments, R⁷ is independently hydrogen. In embodiments, R⁷ isindependently halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂,—CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH,—OCH₃, —OCF₃, —OCHF₂, substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. Inembodiments, R⁷ is independently unsubstituted alkyl, unsubstitutedheteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,unsubstituted aryl, or unsubstituted heteroaryl. In embodiments, R⁷ isindependently substituted or unsubstituted C₁-C₈ alkyl. In embodiments,R⁷ is independently substituted or unsubstituted C₁-C₆ alkyl. Inembodiments, R⁷ is independently substituted or unsubstituted C₁-C₄alkyl. In embodiments, R⁷ is independently unsubstituted C₁-C₈ alkyl. Inembodiments, R⁷ is independently unsubstituted C₁-C₆ alkyl. Inembodiments, R⁷ is independently unsubstituted C₁-C₄ alkyl. Inembodiments, R⁷ is independently unsubstituted methyl.

In embodiments, R² is ═NR⁷. In embodiments, R² is ═NH. In embodiments,R² is ═O. In embodiments, R² is ═S. In embodiments, R⁴ is ═NR⁷. Inembodiments, R⁴ is ═NH. In embodiments, R⁴ is ═O. In embodiments, R⁴ is═S. In embodiments, R² and R⁴ are ═NH. In embodiments, R² and R⁴ are ═O.In embodiments, R² and R⁴ are ═S. In embodiments, R² and R⁴ are ═NR⁷.

In embodiments, L² is a bond. In embodiments, L² is a substituted orunsubstituted alkylene. In embodiments, L² is a substituted orunsubstituted heteroalkylene. In embodiments, L² is L^(2A)-L^(2B)-L^(2C)and L^(2A) is bonded to the substituted or unsubstituted phenyl, whichmay be substituted with R⁵. L^(2A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—. L^(2B) is a bond or substituted or unsubstituted alkylene.L^(2C) is a bond, —O—, or —NH—. In embodiments, L^(2A) is a bond. Inembodiments, L^(2A) is —O—. In embodiments, L^(2A) is —S—. Inembodiments, L^(2A) is —NH—. In embodiments, L^(2A) is —S(O)—. Inembodiments, L^(2A) is —S(O)₂—. In embodiments, L^(2B) is a bond. Inembodiments, L^(2B) is a substituted or unsubstituted alkylene. Inembodiments, L^(2B) is an unsubstituted alkylene. In embodiments, L^(2B)is a substituted or unsubstituted C₁-C₈ alkylene. In embodiments, L^(2B)is an unsubstituted C₁-C₈ alkylene. In embodiments, L^(2B) is asubstituted or unsubstituted C₁-C₆ alkylene. In embodiments, L^(2B) isan unsubstituted C₁-C₆ alkylene. In embodiments, L^(2B) is a substitutedor unsubstituted C₁-C₄ alkylene. In embodiments, L^(2B) is anunsubstituted C₁-C₄ alkylene. In embodiments, L^(2B) is a substitutedalkylene. In embodiments, L^(2B) is a substituted C₁-C₈ alkylene. Inembodiments, L^(2B) is a substituted C₁-C₆ alkylene. In embodiments,L^(2B) is a substituted C₁-C₄ alkylene. In embodiments, L^(2B) is analkylene substituted with —CF₃. In embodiments, L^(2C) is a bond. Inembodiments, L^(2C) is —O—. In embodiments, L^(2C) is —NH—. Inembodiments, L^(2A) is a bond; L^(2B) is unsubstituted methylene; andL²C is —O—.

In embodiments, L⁴ is a bond. In embodiments, L⁴ is a substituted orunsubstituted alkylene. In embodiments, L⁴ is a substituted orunsubstituted heteroalkylene. In embodiments, L⁴ is L^(4A)-L^(4B)-L^(4C)and L^(4A) is bonded to the substituted or unsubstituted phenyl, whichmay be substituted with R⁶. L^(4A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—. L^(4B) is a bond or substituted or unsubstituted alkylene.L^(4C) is a bond, —O—, or —NH—. In embodiments, L^(4A) is a bond. Inembodiments, L^(4A) is —O—. In embodiments, L^(4A) is —S—. Inembodiments, L^(4A) is —NH—. In embodiments, L^(4A) is —S(O)—. Inembodiments, L^(4A) is —S(O)₂—. In embodiments, L^(4B) is a bond. Inembodiments, L^(4B) is a substituted or unsubstituted alkylene. Inembodiments, L^(4B) is an unsubstituted alkylene. In embodiments, L^(4B)is a substituted or unsubstituted C₁-C₈ alkylene. In embodiments, L^(4B)is an unsubstituted C₁-C₄ alkylene. In embodiments, L^(4B) is asubstituted or unsubstituted C₁-C₆ alkylene. In embodiments, L^(4B) isan unsubstituted C₁-C₆ alkylene. In embodiments, L^(4B) is a substitutedor unsubstituted C₁-C₄ alkylene. In embodiments, L^(4B) is anunsubstituted C₁-C₄ alkylene. In embodiments, L^(4B) is a substitutedalkylene. In embodiments, L^(4B) is a substituted C₁-C₄ alkylene. Inembodiments, L^(4B) is a substituted C₁-C₆ alkylene. In embodiments,L^(4B) is a substituted C₁-C₄ alkylene. In embodiments, L^(4B) is analkylene substituted with —CF₃. In embodiments, L^(4C) is a bond. Inembodiments, L^(4C) is —O—. In embodiments, L^(4C) is —NH—. Inembodiments, L^(4A) is a bond; L^(4B) is unsubstituted methylene; andL^(4C) is —O—.

In embodiments, the symbol z2 is 0. In embodiments, the symbol z2 is 1.In embodiments, the symbol z4 is 0. In embodiments, the symbol z4 is 1.In embodiments, the symbols z2 and z4 are 0. In embodiments, the symbolsz2 and z4 are 1. In embodiments, the symbol z5 is 0. In embodiments, thesymbol z5 is 1. In embodiments, the symbol z5 is 2. In embodiments, thesymbol z5 is 3. In embodiments, the symbol z5 is 4. In embodiments, thesymbol z5 is 5. In embodiments, the symbol z6 is 0. In embodiments, thesymbol z6 is 1. In embodiments, the symbol z6 is 2. In embodiments, thesymbol z6 is 3. In embodiments, the symbol z6 is 4. In embodiments, thesymbol z6 is 5.

In embodiment, L¹ is a bond. In embodiment, L¹ is —CH₂—. In embodiment,L¹ is —O—. In embodiment, L¹ is —S—. In embodiment, L¹ is —NH—. Inembodiment, L² is a bond. In embodiment, L² is —CH₂—. In embodiment, L²is —O—. In embodiment, L² is —S—. In embodiment, L² is —NH—. Inembodiment, L³ is —CH₂O—. In embodiment, L³ is —OCH₂—. In embodiment, L³is —CH₂—. In embodiment, L³ is a bond. In embodiment, L³ is —CH₂CH₂—. Inembodiment, L³ is —CH₂CH₂O—. In embodiment, L³ is —OCH2CH2-. Inembodiment, L³ is —CH₂S—. In embodiment, L³ is —SCH₂—. In embodiment, L³is —CH₂S(O)—. In embodiment, L³ is —S(O)CH₂—. In embodiment, L³ is—CH₂S(O)₂—. In embodiment, L³ is —S(O)₂CH₂—. In embodiment, L³ is—CH₂NH—. In embodiment, L³ is —NHCH₂—. In embodiment, L³ is —CH(CH₃)O—.In embodiment, L³ is —OCH(CH₃)—. In embodiment, L³ is —O—. Inembodiment, L³ is —S—. In embodiment, L³ is —NH—. In embodiment, L⁴ is—CH2O-. In embodiment, L⁴ is —OCH₂—. In embodiment, L⁴ is —CH₂—. Inembodiment, L⁴ is a bond. In embodiment, L⁴ is —CH₂CH₂—. In embodiment,L⁴ is —CH₂CH₂O—. In embodiment, L⁴ is —OCH₂CH₂—. In embodiment, L⁴ is—CH₂S—. In embodiment, L⁴ is —SCH₂—. In embodiment, L⁴ is —CH₂S(O)—. Inembodiment, L⁴ is —S(O)CH₂—. In embodiment, L⁴ is —CH₂S(O)₂—. Inembodiment, L⁴ is —S(O)₂CH₂—. In embodiment, L⁴ is —CH₂NH—. Inembodiment, L⁴ is —NHCH₂—. In embodiment, L⁴ is —CH(CH₃)O—. Inembodiment, L⁴ is —OCH(CH₃)—. In embodiment, L⁴ is —O—. In embodiment,L⁴ is —S—. In embodiment, L⁴ is —NH—.

In embodiments, the compound has the formula:

Ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z5, and z6 are asdescribed for compounds of formula (I) above, including embodiments.

In embodiments, the compound has the formula:

L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, z4, z5, and z6 are asdescribed for compounds of formula (I) above, including embodiments. R⁸and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. The symbols b and d areindependently 0 or 1.

In embodiments, R⁸ is hydrogen. In embodiments, R⁸ is substituted orunsubstituted alkyl. In embodiments, R⁸ is substituted or unsubstitutedC₁-C₈ alkyl. In embodiments, R⁸ is substituted or unsubstituted C₁-C₆alkyl. In embodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkyl.In embodiments, R⁸ is substituted or unsubstituted C₁-C₃ alkyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₈ alkenyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkenyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkynyl. Inembodiments, R⁸ is unsubstituted alkyl. In embodiments, R⁸ isunsubstituted C₁-C₈ alkyl. In embodiments, R⁸ is unsubstituted C₁-C₆alkyl. In embodiments, R⁸ is unsubstituted C₁-C₄ alkyl. In embodiments,R⁸ is unsubstituted C₁-C₃ alkyl. In embodiments, R⁸ is unsubstitutedC₁-C₈ alkenyl. In embodiments, R⁸ is unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁸ is unsubstituted C₁-C₄ alkenyl. In embodiments, R⁸ isunsubstituted C₁-C₄ alkynyl. In embodiments, R⁸ is —CCH. In embodiments,R⁸ is

In embodiments, R⁸ is

In embodiments, R⁹ is hydrogen. In embodiments, R⁹ is substituted orunsubstituted alkyl. In embodiments, R⁹ is substituted or unsubstitutedC₁-C₈ alkyl. In embodiments, R⁹ is substituted or unsubstituted C₁-C₆alkyl. In embodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkyl.In embodiments, R⁹ is substituted or unsubstituted C₁-C₃ alkyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₈ alkenyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkenyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkynyl. Inembodiments, R⁹ is unsubstituted alkyl. In embodiments, R⁹ isunsubstituted C₁-C₈ alkyl. In embodiments, R⁹ is unsubstituted C₁-C₆alkyl. In embodiments, R⁹ is unsubstituted C₁-C₄ alkyl. In embodiments,R⁹ is unsubstituted C₁-C₃ alkyl. In embodiments, R⁹ is unsubstitutedC₁-C₈ alkenyl. In embodiments, R⁹ is unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁹ is unsubstituted C₁-C₄ alkenyl. In embodiments, R⁹ isunsubstituted C₁-C₄ alkynyl. In embodiments, R⁹ is —CCH. In embodiments,R⁹ is

In embodiments, R⁹ is

In embodiments, R⁸ and R⁹ are hydrogen.

In embodiments, the symbol b is 0. In embodiments, the symbol b is 1. Inembodiments, the symbol d is 0. In embodiments, the symbol d is 1. Inembodiments, the symbols b and d are 0. In embodiments, the symbols band d are 1. In embodiments, the symbol b is 0 and d is 1. Inembodiments, the symbol b is 1 and d is 0.

In embodiments, the compound has the formula:

L², L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, b, d, z5, and z6 are asdescribed for compounds of formula (I), (Ia), and (III) above, includingembodiments.

In embodiments, the compound has the formula:

L², L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z5, and z6 are as described forcompounds of formula (I), (Ia), (III), and (IIIa) above, includingembodiments.

In embodiments, the compound has the formula:

R^(5.1) and R^(5.2) are as independently described for R⁵, includingembodiments. R^(6.1) and R^(6.2) are as independently described for R⁶,including embodiments. In embodiments, R^(5.1) is independentlyhydrogen, halogen, —CF₃, —CN, —N₃, substituted or unsubstituted C₁-C₄alkyl, substituted or unsubstituted 2 to 4 membered heteroalkyl,substituted or unsubstituted 5 to 6 membered heteroaryl,

In embodiments, R^(6.1) is independently hydrogen, halogen, —CF₃, —CN,—N₃, substituted or unsubstituted C₁-C₄ alkyl, substituted orunsubstituted 2 to 4 membered heteroalkyl, substituted or unsubstituted5 to 6 membered heteroaryl,

In embodiments, R^(5.2) is independently hydrogen, halogen, —CCSi(CH₃)₃,—CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted orunsubstituted 5 to 6 membered heteroaryl,

In embodiments, R^(6.2) is independently hydrogen, halogen, —CCSi(CH₃)₃,—CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted orunsubstituted 5 to 6 membered heteroaryl,

In embodiments, R^(5.1) is independently halogen, unsubstituted C₁-C₃alkyl, or unsubstituted C₁-C₃ haloalkyl. In embodiments, R^(6.1) isindependently halogen, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R⁵² is independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R⁶² is independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R^(5.1) is independently —Cl, —I, —CF₃, —CH₃,or —CCH. In embodiments, R^(6.1) is independently —Cl, —I, —CF₃, —CH₃,or —CCH. In embodiments, R⁵² is independently hydrogen, —Cl, —F, —I,—CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or —CCH. In embodiments, R⁶² isindependently hydrogen, —Cl, —F, —I, —CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or—CCH.

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is ISRIB. In embodiments, the compound istrans-ISRIB. In embodiments, the compound is cis-ISRIB. In embodiments,the compound is a mixture of trans- and cis-ISRIB.In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is a mixture of cis-ISRIB and trans-ISRIB.

In embodiments of the method of treating a disease, the disease isselected from the group consisting of cancer, a neurodegenerativedisease, vanishing white matter disease, childhood ataxia with CNShypo-myelination, and an intellectual disability syndrome. Inembodiments of the method of treating a disease, the disease is cancer.In embodiments of the method of treating a disease, the disease is aneurodegenerative disease. In embodiments of the method of treating adisease, the disease is vanishing white matter disease. In embodimentsof the method of treating a disease, the disease is childhood ataxiawith CNS hypo-myelination. In embodiments of the method of treating adisease, the disease is an intellectual disability syndrome. Inembodiments of the method of treating a disease, the disease isassociated with phosphorylation of eIF2α. In embodiments of the methodof treating a disease, the disease is associated with an eIF2α signalingpathway. In embodiments of the method of treating a disease, the diseaseis a cancer of a secretory cell type. In embodiments of the method oftreating a disease, the disease is pancreatic cancer. In embodiments ofthe method of treating a disease, the disease is breast cancer. Inembodiments of the method of treating a disease, the disease is multiplemyeloma. In embodiments of the method of treating a disease, the diseaseis lymphoma. In embodiments of the method of treating a disease, thedisease is leukemia. In embodiments of the method of treating a disease,the disease is a hematopoietic cell cancer.

In embodiments of the method of treating a disease, the disease isAlzheimer's disease. In embodiments of the method of treating a disease,the disease is Amyotrophic lateral sclerosis. In embodiments of themethod of treating a disease, the disease is Creutzfeldt-Jakob disease.In embodiments of the method of treating a disease, the disease isfrontotemporal dementia. In embodiments of the method of treating adisease, the disease is Gerstmann-Sträussler-Scheinker syndrome. Inembodiments of the method of treating a disease, the disease isHuntington's disease. In embodiments of the method of treating adisease, the disease is HIV-associated dementia. In embodiments of themethod of treating a disease, the disease is kuru. In embodiments of themethod of treating a disease, the disease is Lewy body dementia. Inembodiments of the method of treating a disease, the disease is Multiplesclerosis. In embodiments of the method of treating a disease, thedisease is Parkinson's disease. In embodiments of the method of treatinga disease, the disease is a Prion disease.

In embodiments of the method of treating a disease, the disease is aninflammatory disease. In embodiments, the inflammatory disease ispostoperative cognitive dysfunction. In embodiments, the inflammatorydisease is traumatic brain injury. In embodiments, the inflammatorydisease is arthritis. In embodiments, the inflammatory disease isrheumatoid arthritis. In embodiments, the inflammatory disease ispsoriatic arthritis. In embodiments, the inflammatory disease isjuvenile idiopathic arthritis. In embodiments, the inflammatory diseaseis multiple sclerosis. In embodiments, the inflammatory disease issystemic lupus erythematosus (SLE). In embodiments, the inflammatorydisease is myasthenia gravis. In embodiments, the inflammatory diseaseis juvenile onset diabetes. In embodiments, the inflammatory disease isdiabetes mellitus type 1. In embodiments, the inflammatory disease isGuillain-Barre syndrome. In embodiments, the inflammatory disease isHashimoto's encephalitis. In embodiments, the inflammatory disease isHashimoto's thyroiditis. In embodiments, the inflammatory disease isankylosing spondylitis. In embodiments, the inflammatory disease ispsoriasis. In embodiments, the inflammatory disease is Sjogren'ssyndrome. In embodiments, the inflammatory disease is vasculitis. Inembodiments, the inflammatory disease is glomerulonephritis. Inembodiments, the inflammatory disease is auto-immune thyroiditis. Inembodiments, the inflammatory disease is Behcet's disease. Inembodiments, the inflammatory disease is Crohn's disease. Inembodiments, the inflammatory disease is ulcerative colitis. Inembodiments, the inflammatory disease is bullous pemphigoid. Inembodiments, the inflammatory disease is sarcoidosis. In embodiments,the inflammatory disease is ichthyosis. In embodiments, the inflammatorydisease is Graves ophthalmopathy. In embodiments, the inflammatorydisease is inflammatory bowel disease. In embodiments, the inflammatorydisease is Addison's disease. In embodiments, the inflammatory diseaseis Vitiligo. In embodiments, the inflammatory disease is asthma. Inembodiments, the inflammatory disease is allergic asthma. Inembodiments, the inflammatory disease is acne vulgaris. In embodiments,the inflammatory disease is celiac disease. In embodiments, theinflammatory disease is chronic prostatitis. In embodiments, theinflammatory disease is inflammatory bowel disease. In embodiments, theinflammatory disease is pelvic inflammatory disease. In embodiments, theinflammatory disease is reperfusion injury. In embodiments, theinflammatory disease is sarcoidosis. In embodiments, the inflammatorydisease is transplant rejection. In embodiments, the inflammatorydisease is interstitial cystitis. In embodiments, the inflammatorydisease is atherosclerosis. In embodiments, the inflammatory disease isatopic dermatitis.

In embodiments, the method of treatment is a method of prevention. Forexample, a method of treating postsurgical cognitive dysfunction mayinclude preventing postsurgical cognitive dysfunction or a symptom ofpostsurgical cognitive dysfunction or reducing the severity of a symptomof postsurgical cognitive dysfunction by administering a compounddescribed herein prior to surgery.

In embodiments, the compounds set forth herein are provided aspharmaceutical compositions including the compound and apharmaceutically acceptable excipient. In embodiments of the method, thecompound, or a pharmaceutically acceptable salt thereof, isco-adminstered with a second agent (e.g. therapeutic agent). Inembodiments of the method, the compound, or a pharmaceuticallyacceptable salt thereof, is co-adminstered with a second agent (e.g.therapeutic agent), which is administered in a therapeutically effectiveamount. In embodiments of the method, the second agent is an agent fortreating cancer (e.g. pancreatic cancer, breast cancer, multiplemyeloma, or cancers of secretory cells), neurodegenerative diseases,vanishing white matter disease, childhood ataxia with CNShypo-myelination, and/or intellectual disability syndromes (e.g.associated with impaired function of eIF2 or components in a signaltransduction pathway including eIF2), or an inflammatory disease (e.g.POCD or TBI). In embodiments, the second agent is an anti-cancer agent.In embodiments, the second agent is a chemotherapeutic. In embodiments,the second agent is an agent for improving memory. In embodiments, thesecond agent is an agent for treating a neurodegenerative disease. Inembodiments, the second agent is an agent for treating vanishing whitematter disease. In embodiments, the second agent is an agent fortreating childhood ataxia with CNS hypo-myelination. In embodiments, thesecond agent is an agent for treating an intellectual disabilitysyndrome. In embodiments, the second agent is an agent for treatingpancreatic cancer. In embodiments, the second agent is an agent fortreating breast cancer. In embodiments, the second agent is an agent fortreating multiple myeloma. In embodiments, the second agent is an agentfor treating myeloma. In embodiments, the second agent is an agent fortreating a cancer of a secretory cell. In embodiments, the second agentis an agent for reducing eIF2α phosphorylation. In embodiments, thesecond agent is an agent for inhibiting a pathway activated by eIF2αphosphorylation. In embodiments, the second agent is an agent forinhibiting the integrated stress response. In embodiments, the secondagent is an anti-inflammatory agent.

In some embodiments, the compound is a compound described herein. Insome embodiments, the compound is a compound described in the Examples,an example, a table, the figures, or a figure. In some embodiments, thecompound is a compound described in Table 2. In some embodiments, thecompound is a compound described in the Compounds section below.

The Integrated Stress Response (ISR) is a collection of cellular stressresponse pathways that converge in phosphorylation of the translationinitiation factor eIF2α resulting in a reduction in overall translationin cells. Mammalian cells have four eIF2α kinases that phosphorylatethis initiation factor in the same residue (serine 51); PERK isactivated by the accumulation of unfolded proteins in the endoplasmicreticulum (ER), GCN2 is activated by amino acid starvation, PKR by viralinfection and HRI by heme deficiency. Activation of these kinasesdecreases bulk protein synthesis but it also culminates in increasedexpression of specific mRNAs that contain uORFs. Two examples of thesemRNAs are the transcription factor ATF4 and the pro-apoptotic gene CHOP.Phosphorylation of eIF2α upon stress and the concomitant reduction inprotein translation has been shown to both have cytoprotective andcytotoxic effects depending on the cellular context and duration andseverity of the stress. An integrated stress response-associated diseaseis a disease characterized by increased activity in the integratedstress response (e.g. increased phosphorylation of eIF2α by an eIF2αkinase compared to a control such as a subject without the disease). Adisease associated with phosphorylation of eIF2α is diseasecharacterized by an increase in phosphorylation of eIF2α relative to acontrol, such as a subject without the disease.

Activation of PERK occurs upon ER stress and hypoxic conditions and itsactivation and effect on translation has been shown to be cytoprotectivefor tumor cells [47]. Adaptation to hypoxia in the tumormicroenvironment is critical for survival and metastatic potential. PERKhas also been shown to promote cancer proliferation by limitingoxidative DNA damage and death [48, 49]. Moreover, a newly identifiedPERK inhibitor has been shown to have antitumor activity in a humanpancreatic tumor xenograft mode [50]. Compounds disclosed herein (e.g.ISRIB) decrease the viability of cells that are subjected to ER-stress.Thus, pharmacological and acute inhibition of the PERK branch with thecompounds disclosed herein results in reduced cellular fitness. Duringtumor growth, compounds disclosed herein (e.g. ISRIB), that block thecytoprotective effects of eIF2α phosphorylation upon stress may provepotent anti-proliferative agents.

It is known that under certain stress conditions several eIF2α kinasescan be simultaneously activated. For example, during tumor growth, thelack of nutrients and hypoxic conditions are known to both activate GCN2and PERK. Like PERK, GCN2 and their common target, ATF4, have beenproposed to play a cytoprotective role [51]. By blocking signaling byboth kinases, compounds disclosed herein (e.g. ISRIB) may bypass theability of the ISR to protect cancer cells against the effects of lownutrients and oxygen levels encountered during the growth of the tumor.

Prolonged ER stress leads to the accumulation of CHOP, a pro-apoptoticmolecule. In a prion mouse model, overexpression of the phosphatase ofeIF2α increased survival of prion-infected mice whereas sustained eIF2αphosphorylation decreased survival [52]. The restoration of proteintranslation rates during prion disease was shown to rescue synapticdeficits and neuronal loss. Compounds disclosed herein (e.g. ISRIB) makecells insensitive to eIF2α phosphorylation and thus sustains proteintranslation. Compounds disclosed herein (e.g. ISRIB) could prove potentinhibitors of neuronal cell death in prion disease by blocking thedeleterious effects of prolonged eIF2α phosphorylation. Given theprevalence of protein misfolding and activation on the UPR in severalneurodegenerative diseases (e.g. Alzheimer's (AD) and Parkinson's (PD)),manipulation of the PERK-eIF2α branch could prevent synaptic failure andneuronal death across the spectrum of these disorders.

Another example of tissue-specific pathology that is linked toheightened eIF2α phosphorylation is the fatal brain disorder, vanishingwhite matter disease (VWM) or childhood ataxia with CNS hypo-myelination(CACH). This disease has been linked to mutation in eIF2B, the GTPexchange factor that is necessary for eIF2 function in translation [53].eIF2α phosphorylation inhibits the activity of eIF2B and mutations inthis exchange factor that reduce its exchange activity exacerbate theeffects of eIF2α phosphorylation. The severe consequences of the CACHmutations point to the dangers of UPR hyper-activation, especially as itpertains to the myelin-producing oligodendrocyte. Small molecules, suchcompounds disclosed herein (e.g. ISRIB), that block signaling througheIF2α phosphorylation may reduce the deleterious effects of itshyper-activation in VWM.

Methods of Improving Memory

In another aspect is provided a method of improving long-term memory ina patient, the method including administering a therapeuticallyeffective amount of a compound to the patient, wherein the compound is acompound described herein, including embodiments (e.g. compound offormula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih, II, III, IIIa, IIIb, IIIc, orIV, or any embodiment thereof, including compounds described for use ina different method herein or in the Compounds section below or in anexample, table, figure, or claim). In embodiments, the patient is human.In embodiments, the patient is a non-human mammal. In embodiments, thepatient is a domesticated animal. In embodiments, the patient is a dog.In embodiments, the patient is a bird. In embodiments, the patient is ahorse. In embodiments, the patient is a bovine. In embodiments, thepatient is a primate.

In embodiments, the compounds set forth herein are provided aspharmaceutical compositions including the compound and apharmaceutically acceptable excipient. In embodiments of the method, thecompound, or a pharmaceutically acceptable salt thereof, isco-adminstered with a second agent (e.g. therapeutic agent). Inembodiments of the method, the compound, or a pharmaceuticallyacceptable salt thereof, is co-adminstered with a second agent (e.g.therapeutic agent), which is administered in a therapeutically effectiveamount. In embodiments, the second agent is an agent for improvingmemory.

In some embodiments, the compound is a compound described herein. Insome embodiments, the compound is a compound described in the Examples,an example, a table, the figures, or a figure. In some embodiments, thecompound is a compound described in Table 2.

Induction of long-term memory (LTM) has been shown to be facilitated bydecreased and impaired by increased eIF2α phosphorylation. The datastrongly support the notion that under physiological conditions, adecrease in eIF2α phosphorylation constitutes a critical step for thelong term synaptic changes required for memory formation and ATF4 hasbeen shown to be an important regulator of these processes [54] [55][56]. It is not known what the contributions of the different eIF2αkinases to learning is or whether each play a differential role in thedifferent parts of the brain. Regardless of the eIF2α kinase/sresponsible for phosphorylation of eIF2α in the brain, compoundsdisclosed herein (e.g. ISRIB), block translation attenuation and ATF4production making them ideal molecules to block the effects of thisphosphorylation event on memory. We have shown that pharmacologicaltreatment with compounds disclosed herein (e.g. ISRIB) increases spatialmemory and enhances both auditory and contextual fear conditioning.

Regulators of translation, such as compounds disclosed herein (e.g.ISRIB), could serve as therapeutic agents that improve memory in humandisorders associated with memory loss such as Alzheimer's disease and inother neurological disorders that activate the UPR in neurons and thuscould have negative effects on memory consolidation such as Parkinson'sdisease, Amyotrophic lateral sclerosis and prion diseases. In addition,a mutation in eIF2γ, that disrupts complex integrity linked intellectualdisability (intellectual disability syndrome or ID) to impairedtranslation initiation in humans [57]. Hence, two diseases with impairedeIF2 function, ID and VWM, display distinct phenotypes but both affectmainly the brain and impair learning.

Methods of Increasing Protein Production

We have also shown that compounds disclosed herein (e.g. ISRIB) increasetranslation in an in vitro rabbit reticulocyte translation system.Compounds disclosed herein (e.g. ISRIB) could prove useful inapplications where increasing protein production output is desirable,such as in vitro cell free systems for protein production. In vitrosystems have basal levels of eIF2α phosphorylation that reducetranslational output [58, 59]. Similarly production of antibodies byhybridomas may also be improved by addition of compounds disclosedherein (e.g. ISRIB).

In another aspect is provided a method of increasing protein expressionof a cell or in vitro expression system, the method includingadministering an effective amount of a compound to the cell orexpression system, wherein the compound is a compound described herein,including embodiments (e.g. compound of formula I, Ia, Ib, Ic, Id, Ie,If, Ig, Ih, II, III, IIIa, IIIb, IIIc, or IV, or any embodiment thereof,including compounds described for use in a different method herein or inthe Compounds section below or in an example, table, figure, or claim).In embodiments, the method is a method of increasing protein expressionby a cell and includes administering an effective amount of a compounddescribed herein (e.g. compound of formula I, Ia, Ib, Ic, Id, Ie, If,Ig, Ih, II, III, IIIa, IIIb, IIIc, or IV, or any embodiment thereof,including compounds described for use in a different method herein or inthe Compounds section below or in an example, table, figure, or claim)to the cell. In embodiments, the method is a method of increasingprotein expression by an in vitro protein expression system and includesadministering an effective amount of a compound described herein (e.g.compound of formula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih, II, III, IIIa,IIIb, IIIc, or IV, or any embodiment thereof, including compoundsdescribed for use in a different method herein or in the Compoundssection below or in an example, table, figure, or claim) to the in vitro(e.g. cell free) protein expression system.

In embodiments, the compounds set forth herein are provided aspharmaceutical compositions including the compound and apharmaceutically acceptable excipient. In embodiments of the method, thecompound, or a pharmaceutically acceptable salt thereof, isco-adminstered with a second agent. In embodiments of the method, thecompound, or a pharmaceutically acceptable salt thereof, isco-adminstered with a second agent, which is administered in atherapeutically effective amount. In embodiments, the second agent is anagent for improving protein expression.

In some embodiments, the compound is a compound described herein. Insome embodiments, the compound is a compound described in the Examples,an example, a table, the figures, or a figure. In some embodiments, thecompound is a compound described in Table 2.

Compounds

The compounds described in this Compounds section may be included in anyof the methods described herein. Thus, we have identified a series ofsmall molecule inhibitors (e.g. ISRIB) of the PERK-mediated signal thatleads to translational attenuation in cell-based assays. In addition,the compounds inhibit the action of the other three eIF2α kinases: GCN2,PKR and HRI, which lead to eIF2α phosphorylation on the same residue(serine 51) and thus are ISR inhibitors. The disclosed compounds (e.g.ISRIB), make cells resistant to the effects of eIF2α phosphorylation. Nosmall molecules have been identified that can make cells insensitive tothe effects of eIF2α phosphorylation on translation initiation. To date,these compounds have not shown toxicity and have good Pharmacokineticproperties. These compounds can be used to block translationalregulation by the four eIF2α kinases PERK (activated by ER stress), PKR(activated by viral infection), HRI (activated by heme deficiency) andGCN2 (activated by amino acid starvation).

Compounds useful in the methods disclosed herein are described above andbelow. Thus, the compounds described herein, including those set forthbelow in this Compounds section, are useful in the methods providedhere, including all embodiments thereof. In addition to the compoundsdisclosed above, in another aspect is provided a compound, or apharmaceutically acceptable salt thereof, having the formula:

wherein ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, z4, z5and z6, are as described herein, including embodiments and in the methodof treatment section herein above. In embodiments, Ring A is substitutedor unsubstituted cycloalkylene or substituted or unsubstituted arylene.In embodiments, L¹, L², L³, and L⁴ are independently a bond, —NH—, —O—,—S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene. In embodiments, R¹, R³, R⁵,R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R² and R⁴ areindependently ═NR⁷, ═O, or ═S. The symbols z2 and z4 are eachindependently 0 or 1. The symbols z5 and z6 are each independently aninteger from 0 to 5. In embodiments, the compound is not

In embodiments, ring A is substituted or unsubstituted cycloalkylene. Inembodiments, ring A is substituted or unsubstituted C₃-C₈ cycloalkylene.In embodiments, ring A is substituted or unsubstituted C₃-C₆cycloalkylene. In embodiments, ring A is substituted or unsubstitutedC₃-C₄ cycloalkylene. In embodiments, ring A is substituted orunsubstituted C₄-C₈ cycloalkylene. In embodiments, ring A is substitutedor unsubstituted C₄-C₆ cycloalkylene. In embodiments, ring A issubstituted or unsubstituted cyclohexylene. In embodiments, ring A issubstituted or unsubstituted cyclobutylene. In embodiments, ring A issubstituted or unsubstituted cyclopentylene. In embodiments, ring A issubstituted or unsubstituted C₄-C₆ cycloalkenylene. In embodiments, ringA is unsubstituted cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₈ cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₆ cycloalkylene. In embodiments, ring A isunsubstituted C₃-C₄ cycloalkylene. In embodiments, ring A isunsubstituted C₄-C₈ cycloalkylene. In embodiments, ring A isunsubstituted C₄-C₆ cycloalkylene. In embodiments, ring A isunsubstituted cyclohexylene. In embodiments, ring A is unsubstitutedcyclobutylene. In embodiments, ring A is unsubstituted cyclopentylene.In embodiments, ring A is unsubstituted C₄-C₆ cycloalkenylene. Inembodiments, ring A is substituted or unsubstituted arylene. Inembodiments, ring A is substituted or unsubstituted C₆-C₁₀ arylene. Inembodiments, ring A is substituted or unsubstituted phenylene. Inembodiments, ring A is substituted or unsubstituted naphthylene. Inembodiments, ring A is unsubstituted C₆-C₁₀ arylene. In embodiments,ring A is unsubstituted phenylene. In embodiments, ring A isunsubstituted naphthylene.

L¹ may be a bond or substituted or unsubstituted alkylene. L¹ may besubstituted or unsubstituted C₁-C₅ alkylene. L¹ may be substituted orunsubstituted C₁-C₃ alkylene. L¹ may be substituted or unsubstitutedmethylene. L¹ may be a bond. L¹ may be an unsubstituted alkylene. L¹ maybe an unsubstituted methylene. L¹ may be an unsubstituted ethylene. L¹may be a methylene substituted with an unsubstituted alkyl L¹ may be amethylene substituted with an unsubstituted C₁-C₄ alkyl L¹ may be amethylene substituted with an unsubstituted C₁-C₃ alkyl.

L³ may be a bond or substituted or unsubstituted alkylene. L³ may besubstituted or unsubstituted C₁-C₅ alkylene. L³ may be substituted orunsubstituted C₁-C₃ alkylene. L³ may be substituted or unsubstitutedmethylene. L³ may be a bond. L³ may be an unsubstituted alkylene. L³ maybe an unsubstituted methylene. L³ may be an unsubstituted ethylene. L³may be a methylene substituted with an unsubstituted alkyl L³ may be amethylene substituted with an unsubstituted C₁-C₄ alkyl L³ may be amethylene substituted with an unsubstituted C₁-C₃ alkyl. In embodiments,L¹ and L³ may be a bond. In embodiments, L¹ and L³ may independently bean unsubstituted alkylene. In embodiments, L¹ and L³ may be anunsubstituted methylene.

In embodiments, R¹ is hydrogen. In embodiments, R¹ is —CH₂CCH. Inembodiments, R¹ is

In embodiments, R¹ is

In embodiments, R¹ is

In embodiments, R¹ is substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. Inembodiments, R¹ is substituted or unsubstituted alkyl. In embodiments,R¹ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R¹ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R¹ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R¹ isunsubstituted alkyl. In embodiments, R¹ is unsubstituted C₁-C₈ alkyl. Inembodiments, R¹ is unsubstituted C₁-C₆ alkyl. In embodiments, R¹ isunsubstituted C₁-C₄ alkyl. In embodiments, R¹ is substituted orunsubstituted heteroalkyl. In embodiments, R¹ is substituted orunsubstituted 2 to 8 membered heteroalkyl. In embodiments, le isunsubstituted 2 to 8 membered heteroalkyl.

In embodiments, R³ is hydrogen. In embodiments, R³ is —CH₂CCH. Inembodiments, R³ is

In embodiments, R³ is

In embodiments, R³ is

In embodiments, R³ is substituted or unsubstituted alkyl, substituted orunsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl,substituted or unsubstituted heterocycloalkyl, substituted orunsubstituted aryl, or substituted or unsubstituted heteroaryl. Inembodiments, R³ is substituted or unsubstituted alkyl. In embodiments,R³ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R³ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R³ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R³ isunsubstituted alkyl. In embodiments, R³ is unsubstituted C₁-C₈ alkyl. Inembodiments, R³ is unsubstituted C₁-C₆ alkyl. In embodiments, R³ isunsubstituted C₁-C₄ alkyl. In embodiments, R³ is substituted orunsubstituted heteroalkyl. In embodiments, R³ is substituted orunsubstituted 2 to 8 membered heteroalkyl. In embodiments, R³ isunsubstituted 2 to 8 membered heteroalkyl.

In embodiments, R⁵ is independently halogen, —OCH₃, —OCH₂Ph, —C(O)Ph,—CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂,—C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H,—SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R⁵ isindependentlyhalogen, —OCH₃, —OCH₂Ph, —CH₃, —OH, —CF₃, —CCl₃, —CN, —S(O)CH₃, —NO₂,—C(O)CH₃, —C(O)Ph, —CH(CH₃)₂, —CCSi(CH₃)₃, or —CCH. In embodiments, R⁵is independently halogen. In embodiments, R⁵ is independently —OCH₃. Inembodiments, R⁵ is independently —OCH₂Ph. In embodiments, R⁵ isindependently —CH₃. In embodiments, R⁵ is independently —OH. Inembodiments, R⁵ is independently —CF₃. In embodiments, R⁵ isindependently —CCl₃. In embodiments, R⁵ is independently —CN. Inembodiments, R⁵ is independently —S(O)CH₃. In embodiments, R⁵ isindependently —NO₂. In embodiments, R⁵ is independently —C(O)CH₃. Inembodiments, R⁵ is independently —C(O)Ph. In embodiments, R⁵ isindependently —CH(CH₃)₂. In embodiments, R⁵ is independently—CCSi(CH₃)₃. In embodiments, R⁵ is independently —CCH. In embodiments,R⁵ is —F. In embodiments, R⁵ is —Cl. In embodiments, R⁵ is —Br. Inembodiments, R⁵ is —I. In embodiments, R⁵ is substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. In embodiments, R⁵ is unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. Inembodiments, R⁵ is substituted or unsubstituted alkyl. In embodiments,R⁵ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R⁵ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R⁵ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R⁵ isunsubstituted alkyl. In embodiments, R⁵ is unsubstituted C₁-C₈ alkyl. Inembodiments, R⁵ is unsubstituted C₁-C₆ alkyl. In embodiments, R⁵ isunsubstituted C₁-C₄ alkyl. In embodiments, R⁵ is substituted alkyl. Inembodiments, R⁵ is substituted C₁-C₈ alkyl. In embodiments, R⁵ issubstituted C₁-C₆ alkyl. In embodiments, R⁵ is substituted C₁-C₄ alkyl.In embodiments, R⁵ is substituted C₁-C₃ alkyl. In embodiments, R⁵ issubstituted or unsubstituted heteroalkyl. In embodiments, R⁵ issubstituted or unsubstituted 2 to 8 membered heteroalkyl. Inembodiments, R⁵ is unsubstituted 2 to 6 membered heteroalkyl. Inembodiments, R⁵ is unsubstituted 2 to 4 membered heteroalkyl. Inembodiments, R⁵ is substituted 2 to 8 membered heteroalkyl. Inembodiments, R⁵ is substituted 2 to 6 membered heteroalkyl. Inembodiments, R⁵ is substituted 2 to 4 membered heteroalkyl. Inembodiments, R⁵ is independently —N₃. In embodiments, R⁵ isindependently —C(NN)CF₃. In embodiments, R⁵ is independently—C(NH—NH)CF₃. In embodiments, R⁵ is

In embodiments, R⁶ is independently halogen, —OCH₃, —OCH₂Ph, —C(O)Ph,—CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂,—C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H,—SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R⁶ isindependentlyhalogen, —OCH₃, —OCH₂Ph, —CH₃, —OH, —CF₃, —CCl₃, —CN, —S(O)CH₃, —NO₂,—C(O)CH₃, —C(O)Ph, —CH(CH₃)₂, —CCSi(CH₃)₃, or —CCH. In embodiments, R⁶is independently halogen. In embodiments, R⁶ is independently —OCH₃. Inembodiments, R⁶ is independently —OCH₂Ph. In embodiments, R⁶ isindependently —CH₃. In embodiments, R⁶ is independently —OH. Inembodiments, R⁶ is independently —CF₃. In embodiments, R⁶ isindependently —CCl₃. In embodiments, R⁶ is independently —CN. Inembodiments, R⁶ is independently —S(O)CH₃. In embodiments, R⁶ isindependently —NO₂. In embodiments, R⁶ is independently —C(O)CH₃. Inembodiments, R⁶ is independently —C(O)Ph. In embodiments, R⁶ isindependently —CH(CH₃)₂. In embodiments, R⁶ is independently—CCSi(CH₃)₃. In embodiments, R⁶ is independently —CCH. In embodiments,R⁶ is —F. In embodiments, R⁶ is —Cl. In embodiments, R⁶ is —Br. Inembodiments, R⁶ is —I. In embodiments, R⁶ is substituted orunsubstituted alkyl, substituted or unsubstituted heteroalkyl,substituted or unsubstituted cycloalkyl, substituted or unsubstitutedheterocycloalkyl, substituted or unsubstituted aryl, or substituted orunsubstituted heteroaryl. In embodiments, R⁶ is unsubstituted alkyl,unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstitutedheterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. Inembodiments, R⁶ is substituted or unsubstituted alkyl. In embodiments,R⁶ is substituted or unsubstituted C₁-C₈ alkyl. In embodiments, R⁶ issubstituted or unsubstituted C₁-C₆ alkyl. In embodiments, R⁶ issubstituted or unsubstituted C₁-C₄ alkyl. In embodiments, R⁶ isunsubstituted alkyl. In embodiments, R⁶ is unsubstituted C₁-C₈ alkyl. Inembodiments, R⁶ is unsubstituted C₁-C₆ alkyl.

In embodiments, R⁶ is unsubstituted C₁-C₄ alkyl. In embodiments, R⁶ issubstituted alkyl. In embodiments, R⁶ is substituted C₁-C₈ alkyl. Inembodiments, R⁶ is substituted C₁-C₆ alkyl. In embodiments, R⁶ issubstituted C₁-C₄ alkyl. In embodiments, R⁶ is substituted C₁-C₃ alkyl.In embodiments, R⁶ is substituted or unsubstituted heteroalkyl. Inembodiments, R⁶ is substituted or unsubstituted 2 to 8 memberedheteroalkyl. In embodiments, R⁶ is unsubstituted 2 to 6 memberedheteroalkyl. In embodiments, R⁶ is unsubstituted 2 to 4 memberedheteroalkyl. In embodiments, R⁶ is substituted 2 to 8 memberedheteroalkyl. In embodiments, R⁶ is substituted 2 to 6 memberedheteroalkyl. In embodiments, R⁶ is substituted 2 to 4 memberedheteroalkyl. In embodiments, R⁶ is independently —N₃. In embodiments, R⁶is independently —C(NN)CF₃. In embodiments, R⁶ is independently—C(NH—NH)CF₃. In embodiments, R⁶ is

In embodiments, R⁷ is independently hydrogen. In embodiments, R⁷ isindependently halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂,—CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. In embodiments, R⁷ isindependently unsubstituted alkyl, unsubstituted heteroalkyl,unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstitutedaryl, or unsubstituted heteroaryl. In embodiments, R⁷ is independentlysubstituted or unsubstituted C₁-C₈ alkyl. In embodiments, R⁷ isindependently substituted or unsubstituted C₁-C₆ alkyl. In embodiments,R⁷ is independently substituted or unsubstituted C₁-C₄ alkyl. Inembodiments, R⁷ is independently unsubstituted C₁-C₈ alkyl. Inembodiments, R⁷ is independently unsubstituted C₁-C₆ alkyl. Inembodiments, R⁷ is independently unsubstituted C₁-C₄ alkyl. Inembodiments, R⁷ is independently unsubstituted methyl.

In embodiments, R² is ═NR⁷. In embodiments, R² is ═NH. In embodiments,R² is ═O. In embodiments, R² is ═S. In embodiments, R⁴ is ═NR⁷. Inembodiments, R⁴ is ═NH. In embodiments, R⁴ is ═O. In embodiments, R⁴ is═S. In embodiments, R² and R⁴ are ═NH. In embodiments, R² and R⁴ are ═O.In embodiments, R² and R⁴ are ═S. In embodiments, R² and R⁴ are ═NR⁷.

In embodiments, L² is a bond. In embodiments, L² is a substituted orunsubstituted alkylene. In embodiments, L² is a substituted orunsubstituted heteroalkylene. In embodiments, L² is L^(2A)-L^(2B)-L^(2C)and L^(2A) is bonded to the substituted or unsubstituted phenyl, whichmay be substituted with R⁵. L^(2A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—. L^(2B) is a bond or substituted or unsubstituted alkylene. L²Cis a bond, —O—, or —NH—. In embodiments, L^(2A) is a bond. Inembodiments, L^(2A) is —O—. In embodiments, L^(2A) is —S—. Inembodiments, L^(2A) is —NH—. In embodiments, L^(2A) is —S(O)—. Inembodiments, L^(2A) is —S(O)₂—. In embodiments, L^(2B) is a bond. Inembodiments, L^(2B) is a substituted or unsubstituted alkylene. Inembodiments, L^(2B) is an unsubstituted alkylene. In embodiments, L^(2B)is a substituted or unsubstituted C₁—C alkylene. In embodiments, L^(2B)is an unsubstituted C₁-C₈ alkylene. In embodiments, L^(2B) is asubstituted or unsubstituted C₁-C₆ alkylene. In embodiments, L^(2B) isan unsubstituted C₁-C₆ alkylene. In embodiments, L^(2B) is a substitutedor unsubstituted C₁-C₄ alkylene. In embodiments, L^(2B) is anunsubstituted C₁-C₄ alkylene. In embodiments, L^(2B) is a substitutedalkylene. In embodiments, L^(2B) is a substituted C₁-C₆ alkylene. Inembodiments, L^(2B) is a substituted C₁-C₆ alkylene. In embodiments,L^(2B) is a substituted C₁-C₄ alkylene. In embodiments, L^(2B) is analkylene substituted with —CF₃. In embodiments, L^(2C) is a bond. Inembodiments, L^(2C) is —O—. In embodiments, L^(2C) is —NH—. Inembodiments, L^(2A) is a bond; L^(2B) is unsubstituted methylene; andL^(2C) is —O—.

In embodiments, L⁴ is a bond. In embodiments, L⁴ is a substituted orunsubstituted alkylene. In embodiments, L⁴ is a substituted orunsubstituted heteroalkylene. In embodiments, L⁴ is L^(4A)-L^(4B)-L^(4C)and L^(4A) is bonded to the substituted or unsubstituted phenyl, whichmay be substituted with R⁶. L^(4A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—. L^(4B) is a bond or substituted or unsubstituted alkylene.L^(4c) is a bond, —O—, or —NH—. In embodiments, L^(4A) is a bond. Inembodiments, L^(4A) is —O—. In embodiments, L^(4A) is —S—. Inembodiments, L^(4A) is —NH—. In embodiments, L^(4A) is —S(O)—. Inembodiments, L^(4A) is —S(O)₂—. In embodiments, L^(4B) is a bond. Inembodiments, L^(4B) is a substituted or unsubstituted alkylene. Inembodiments, L^(4B) is an unsubstituted alkylene. In embodiments, L^(4B)is a substituted or unsubstituted C₁—C alkylene. In embodiments, L^(4B)is an unsubstituted C₁-C₈ alkylene. In embodiments, L^(4B) is asubstituted or unsubstituted C₁-C₆ alkylene. In embodiments, L^(4B) isan unsubstituted C₁-C₆ alkylene. In embodiments, L^(4B) is a substitutedor unsubstituted C₁-C₄ alkylene. In embodiments, L^(4B) is anunsubstituted C₁-C₄ alkylene. In embodiments, L^(4B) is a substitutedalkylene. In embodiments, L^(4B) is a substituted C₁-C₈ alkylene. Inembodiments, L^(4B) is a substituted C₁-C₆ alkylene. In embodiments,L^(4B) is a substituted C₁-C₄ alkylene. In embodiments, L^(4B) is analkylene substituted with —CF₃. In embodiments, L^(4C) is a bond. Inembodiments, L^(4C) is —O—. In embodiments, L^(4C) is —NH—. Inembodiments, L^(4A) is a bond; L^(4B) is unsubstituted methylene; andL^(4C) is —O—.

In embodiments, the symbol z2 is 0. In embodiments, the symbol z2 is 1.In embodiments, the symbol z4 is 0. In embodiments, the symbol z4 is 1.In embodiments, the symbols z2 and z4 are 0. In embodiments, the symbolsz2 and z4 are 1. In embodiments, the symbol z5 is 0. In embodiments, thesymbol z5 is 1. In embodiments, the symbol z5 is 2. In embodiments, thesymbol z5 is 3. In embodiments, the symbol z5 is 4. In embodiments, thesymbol z5 is 5. In embodiments, the symbol z6 is 0. In embodiments, thesymbol z6 is 1. In embodiments, the symbol z6 is 2. In embodiments, thesymbol z6 is 3. In embodiments, the symbol z6 is 4. In embodiments, thesymbol z6 is 5.

In embodiment, L is a bond. In embodiment, L¹ is —CH₂—. In embodiment,L¹ is —O—. In embodiment, L is —S—. In embodiment, L¹ is —NH—. Inembodiment, L² is a bond. In embodiment, L² is —CH₂—. In embodiment, L²is —O—. In embodiment, L² is —S—. In embodiment, L² is —NH—. Inembodiment, L³ is —CH₂O—. In embodiment, L³ is —OCH₂—. In embodiment, L³is —CH₂—. In embodiment, L³ is a bond. In embodiment, L³ is —CH₂CH₂—. Inembodiment, L³ is —CH₂CH₂O—. In embodiment, L³ is —OCH2CH2-. Inembodiment, L³ is —CH₂S—. In embodiment, L³ is —SCH₂—. In embodiment, L³is —CH₂S(O)—. In embodiment, L³ is —S(O)CH₂—. In embodiment, L³ is—CH₂S(O)₂—. In embodiment, L³ is —S(O)₂CH₂—. In embodiment, L³ is—CH₂NH—. In embodiment, L³ is —NHCH₂—. In embodiment, L³ is —CH(CH₃)O—.In embodiment, L³ is —OCH(CH₃)—. In embodiment, L³ is —O—. Inembodiment, L³ is —S—. In embodiment, L³ is —NH—. In embodiment, L⁴ is—CH₂O—. In embodiment, L⁴ is —OCH₂—. In embodiment, L⁴ is —CH₂—. Inembodiment, L⁴ is a bond. In embodiment, L⁴ is —CH₂CH₂—. In embodiment,L⁴ is —CH₂CH₂O—. In embodiment, L⁴ is —OCH₂CH₂—. In embodiment, L⁴ is—CH₂S—. In embodiment, L⁴ is —SCH₂—. In embodiment, L⁴ is —CH₂S(O)—. Inembodiment, L⁴ is —S(O)CH₂—. In embodiment, L⁴ is —CH₂S(O)₂—. Inembodiment, L⁴ is —S(O)₂CH₂—. In embodiment, L⁴ is —CH₂NH—. Inembodiment, L⁴ is —NHCH₂—. In embodiment, L⁴ is —CH(CH₃)O—. Inembodiment, L⁴ is —OCH(CH₃)—. In embodiment, L⁴ is —O—. In embodiment,L⁴ is —S—. In embodiment, L⁴ is —NH—.

In embodiments, the compound has the formula:

Ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z5, and z6 are asdescribed for compounds of formula (I) above, including embodiments.

In embodiments, the compound has the formula:

L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, z4, z5, and z6 are asdescribed for compounds of formula (I) above, including embodiments. R⁸and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl. The symbols b and d areindependently 0 or 1.

In embodiments, R⁸ is hydrogen. In embodiments, R⁸ is substituted orunsubstituted alkyl. In embodiments, R⁸ is substituted or unsubstitutedC₁-C₈ alkyl. In embodiments, R⁸ is substituted or unsubstituted C₁-C₆alkyl. In embodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkyl.In embodiments, R⁸ is substituted or unsubstituted C₁-C₃ alkyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₈ alkenyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkenyl. Inembodiments, R⁸ is substituted or unsubstituted C₁-C₄ alkynyl. Inembodiments, R⁸ is unsubstituted alkyl. In embodiments, R⁸ isunsubstituted C₁-C₈ alkyl. In embodiments, R⁸ is unsubstituted C₁-C₆alkyl. In embodiments, R⁸ is unsubstituted C₁-C₄ alkyl. In embodiments,R is unsubstituted C₁-C₃ alkyl. In embodiments, R⁸ is unsubstitutedC₁-C₈ alkenyl. In embodiments, R⁸ is unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁸ is unsubstituted C₁-C₄ alkenyl. In embodiments, R⁸ isunsubstituted C₁-C₄ alkynyl. In embodiments, R⁸ is —CCH. In embodiments,R⁸ is

In embodiments, R is

In embodiments, R⁹ is hydrogen. In embodiments, R⁹ is substituted orunsubstituted alkyl. In embodiments, R⁹ is substituted or unsubstitutedC₁-C₈ alkyl. In embodiments, R⁹ is substituted or unsubstituted C₁-C₆alkyl. In embodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkyl.In embodiments, R⁹ is substituted or unsubstituted C₁-C₃ alkyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₈ alkenyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkenyl. Inembodiments, R⁹ is substituted or unsubstituted C₁-C₄ alkynyl. Inembodiments, R⁹ is unsubstituted alkyl. In embodiments, R⁹ isunsubstituted C₁-C₈ alkyl. In embodiments, R⁹ is unsubstituted C₁-C₆alkyl. In embodiments, R⁹ is unsubstituted C₁-C₄ alkyl. In embodiments,R⁹ is unsubstituted C₁-C₃ alkyl. In embodiments, R⁹ is unsubstitutedC₁-C₈ alkenyl. In embodiments, R⁹ is unsubstituted C₁-C₈ alkynyl. Inembodiments, R⁹ is unsubstituted C₁-C₄ alkenyl. In embodiments, R⁹ isunsubstituted C₁-C₄ alkynyl. In embodiments, R⁹ is —CCH. In embodiments,R⁹ is

In embodiments, R⁹ is

In embodiments, R⁸ and R⁹ are hydrogen.

In embodiments, the symbol b is 0. In embodiments, the symbol b is 1. Inembodiments, the symbol d is 0. In embodiments, the symbol d is 1. Inembodiments, the symbols b and d are 0. In embodiments, the symbols band d are 1. In embodiments, the symbol b is 0 and d is 1. Inembodiments, the symbol b is 1 and d is 0.

In embodiments, the compound has the formula:

L², L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, b, d, z5, and z6 are asdescribed for compounds of formula (I), (Ia), and (III) above, includingembodiments.

In embodiments, the compound has the formula:

L², L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z5, and z6 are as described forcompounds of formula (I), (Ia), (III), and (IIIa) above, includingembodiments.

In embodiments, the compound has the formula:

R^(5.1) and R^(5.2) are independently as described for R⁵, includingembodiments. R^(6.1) and R^(6.2) are independently as described for R⁶,including embodiments. In embodiments, R^(5.1) is independentlyhydrogen, halogen, —CF₃, —CN, —N₃, substituted or unsubstituted C₁-C₄alkyl, substituted or unsubstituted 2 to 4 membered heteroalkyl,substituted or unsubstituted 5 to 6

In embodiments, R^(6.1) is independently hydrogen, halogen, —CF₃, —CN,—N₃, substituted or unsubstituted C₁-C₄ alkyl, substituted orunsubstituted 2 to 4 membered heteroalkyl, substituted or unsubstituted5 to 6

In embodiments, R^(5.2) is independently hydrogen, halogen, —CCSi(CH₃)₃,—CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted orunsubstituted 5 to 6 membered heteroaryl,

In embodiments, R^(6.2) is independently hydrogen, halogen, —CCSi(CH₃)₃,—CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted orunsubstituted 5 to 6 membered heteroaryl,

In embodiments, R^(5.1) is independently halogen, unsubstituted C₁-C₃alkyl, or unsubstituted C₁-C₃ haloalkyl. In embodiments, R^(6.1) isindependently halogen, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R^(5.2) is independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R^(6.2) is independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl. In embodiments, R^(5.1) is independently —Cl, —I, —CF₃, —CH₃,or —CCH. In embodiments, R^(6.1) is independently —Cl, —I, —CF₃, —CH₃,or —CCH. In embodiments, R^(5.2) is independently hydrogen, —C, —F, —I,—CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or —CCH. In embodiments, R^(6.2) isindependently hydrogen, —Cl, —F, —I, —CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or—CCH. In embodiments, R^(5.1) is independently hydrogen. In embodiments,R^(5.1) is independently halogen. In embodiments, R^(5.1) isindependently —CF₃. In embodiments, R^(5.1) is independently —CN. Inembodiments, R^(5.1) is independently —N₃. In embodiments, R^(5.1) isindependently unsubstituted C₁-C₄ alkyl. In embodiments, R^(5.1) isindependently unsubstituted 2 to 4 membered heteroalkyl. In embodiments,R^(5.1) is independently unsubstituted 5 to 6 membered heteroaryl. Inembodiments, R^(5.1) is independently substituted C₁-C₄ alkyl. Inembodiments, R^(5.1) is independently substituted 2 to 4 memberedheteroalkyl. In embodiments, R^(5.1) is independently substituted 5 to 6membered heteroaryl. In embodiments, R^(6.1) is independently hydrogen.In embodiments, R^(6.1) is independently halogen. In embodiments,R^(6.1) is independently —CF₃. In embodiments, R^(6.1) is independently—CN. In embodiments, R^(6.1) is independently —N₃. In embodiments,R^(6.1) is independently unsubstituted C₁-C₄ alkyl. In embodiments,R^(6.1) is independently unsubstituted 2 to 4 membered heteroalkyl. Inembodiments, R^(6.1) is independently unsubstituted 5 to 6 memberedheteroaryl. In embodiments, R^(6.1) is independently substituted C₁-C₄alkyl. In embodiments, R^(6.1) is independently substituted 2 to 4membered heteroalkyl. In embodiments, R^(6.1) is independentlysubstituted 5 to 6 membered heteroaryl. In embodiments, R^(5.2) isindependently hydrogen. In embodiments, R^(5.2) is independentlyhalogen. In embodiments, R^(5.2) is independently —CCSi(CH₃)₃. Inembodiments, R^(5.2) is independently —CF₃. In embodiments, R^(5.2) isindependently —NO₂. In embodiments, R^(5.2) is independently —CN. Inembodiments, R^(5.2) is independently —N₃. In embodiments, R^(5.2) isindependently unsubstituted C₁-C₄ alkyl. In embodiments, R^(5.2) isindependently unsubstituted 2 to 4 membered heteroalkyl. In embodiments,R^(5.2) is independently unsubstituted 5 to 6 membered heteroaryl. Inembodiments, R^(5.2) is independently substituted C₁-C₄ alkyl. Inembodiments, R^(5.2) is independently substituted 2 to 4 memberedheteroalkyl. In embodiments, R^(5.2) is independently substituted 5 to 6membered heteroaryl. In embodiments, R^(6.2) is independently hydrogen.In embodiments, R^(6.2) is independently halogen. In embodiments,R^(6.2) is independently —CCSi(CH₃)₃. In embodiments, R^(6.2) isindependently —CF₃. In embodiments, R^(6.2) is independently —NO₂. Inembodiments, R^(6.2) is independently —CN. In embodiments, R^(6.2) isindependently —N₃. In embodiments, R^(6.2) is independentlyunsubstituted C₁-C₄ alkyl. In embodiments, R^(6.2) is independentlyunsubstituted 2 to 4 membered heteroalkyl. In embodiments, R^(6.2) isindependently unsubstituted 5 to 6 membered heteroaryl. In embodiments,R^(6.2) is independently substituted C₁-C₄ alkyl. In embodiments,R^(6.2) is independently substituted 2 to 4 membered heteroalkyl. Inembodiments, R^(6.2) is independently substituted 5 to 6 memberedheteroaryl.

In embodiments, the compound has the formula:

wherein R^(5.1) and R^(5.2) are independently as described for R⁵,including embodiments. R^(6.1) and R^(6.2) are independently asdescribed for R⁶, including embodiments.

In embodiments, the compound has the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, and z4,are as described herein. In embodiments, the compound has the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, and z4,are as described herein. In embodiments, the compound has the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, z2, and z4,are as described herein. In embodiments, the compound has the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁵, z2, and z4, are asdescribed herein. R^(5.1) and R^(5.2) are independently as described forR⁵, including embodiments. R^(6.1) and R^(6.2) are independently asdescribed for R⁶, including embodiments. In embodiments, the compoundhas the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁷, z2, and z4, are asdescribed herein. R^(5.1) and R^(5.2) are independently as described forR⁵, including embodiments. R^(6.1) and R^(6.2) are independently asdescribed for R⁶, including embodiments. In embodiments, the compoundhas the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁷, z2, and z4, are asdescribed herein. R^(5.1), R^(5.2), and R⁵³ are independently asdescribed for R⁵, including embodiments. R^(6.1), R^(6.2), and R^(6.3)are independently as described for R⁶, including embodiments. Inembodiments, the compound has the formula:

wherein, ring A, L¹, L², L³, L⁴, R¹, R², R³, R⁴, R⁷, z2, and z4, are asdescribed herein. R^(5.1), R^(5.2), and R⁵³ are independently asdescribed for R⁵, including embodiments. R^(6.1), R^(6.2), and R^(6.3)are independently as described for R⁶, including embodiments.

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is ISRIB. In embodiments, the compound istrans-ISRIB. In embodiments, the compound is cis-ISRIB. In embodiments,the compound is a mixture of trans- and cis-ISRIB.In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is

In embodiments, the compound is a mixture of cis-ISRIB and trans-ISRIB.

In embodiments, the compound is not

wherein L¹ and L² are both a bond or an unsubstituted C₁-C₂ alkylene. Inembodiments, the compound is not a compound of formula Va wherein L¹ andL² are both a bond or an unsubstituted C₁-C₃ alkylene. In embodiments,the compound is not a compound of formula Va wherein L¹ and L² are botha bond or an unsubstituted C₁-C₄ alkylene.

In embodiments, the compound is not

wherein L³ and L⁴ are both unsubstituted C₁-C₂ alkylene or unsubstituted2 to 3 membered heteroalkylene. In embodiments, the compound is not acompound of formula Vb wherein L³ and L⁴ are both unsubstituted C₁-C₃alkylene or unsubstituted 2 to 3 membered heteroalkylene. Inembodiments, the compound is not a compound of formula Vb wherein L³ andL⁴ are both unsubstituted C₁-C₃ alkylene or unsubstituted 2 to 4membered heteroalkylene.

In embodiments, the compound is not

wherein L¹ and L² are both a bond or an unsubstituted C₁-C₂ alkylene andL³ and L⁴ are both unsubstituted C₁-C₂ alkylene or unsubstituted 2 to 3membered heteroalkylene, and X^(b) is —Cl. In embodiments, the compoundis not a compound of formula (Vc) wherein L¹ and L² are both a bond oran unsubstituted C₁-C₂ alkylene and L³ and L⁴ are both unsubstitutedC₁-C₂ alkylene or unsubstituted 2 to 3 membered heteroalkylene, andX^(b) is a halide. In embodiments, the compound is not a compound offormula (Vc) wherein L¹ and L² are both a bond or an unsubstituted C₁-C₃alkylene and L³ and L⁴ are both unsubstituted C₁-C₃ alkylene orunsubstituted 2 to 3 membered heteroalkylene, and X^(b) is a halide. Inembodiments, the compound is not a compound of formula (Vc) wherein L¹and L² are both a bond or an unsubstituted C₁-C₃ alkylene and L³ and L⁴are both unsubstituted C₁-C₃ alkylene or unsubstituted 2 to 4 memberedheteroalkylene, and X^(b) is a halide.

In embodiments, the compound is not

wherein L¹ and L² are both a bond or an unsubstituted methylene andX^(b) is a halide. In embodiments, the compound is not a compound offormula (Vd) wherein L¹ and L² are independently a bond or anunsubstituted methylene and X^(b) is a halide.

In embodiments, the compound is not

wherein L¹ and L² are both a bond or an unsubstituted methylene. Inembodiments, the compound is not a compound of formula (Ve) wherein L¹and L² are independently a bond or an unsubstituted methylene.

In embodiments, the compound is not

In embodiments, the compound is not

In embodiments, the compound is not

wherein X^(c) is a halide. In embodiments, the compound is not acompound of formula (VIc) wherein X^(c) is halide or —CH₃. Inembodiments, the compound is not a compound of formula (VIc) whereinX^(c) is halide, —CH₃, or —CF₃. In embodiments, the compound is not acompound of formula (VIc) wherein X^(c) is halide, —CH₃, —CCl₃, or —CF₃.In embodiments, the compound is not a compound of formula (VIc) whereinX^(c) is halide, —CH₃, —CCl₃, —CN, or —CF₃. In embodiments, the compoundis not a compound of formula (VIc) wherein X^(c) is halide, —CH₃, —CCl₃,—OH, —CN, or —CF₃. In embodiments, the compound is not a compound offormula (VIc) wherein X^(c) is halide, —CH₃, —CCl₃, —OH, —SH, —CN,—OCH₃, or —CF₃. In embodiments, the compound is not a compound offormula (VIc) wherein X^(c) is unsubstituted C₁-C₂ alkyl, halide, CCl₃,—OH, —SH, —CN, —OCH₃, or —CF₃. In embodiments, the compound is not acompound of formula (VIc) wherein X^(c) is unsubstituted C₁-C₃ alkyl,halide, CCl₃, —OH, —SH, —CN, —OCH₃, or —CF₃. In embodiments, thecompound is not a compound of formula (VIc) wherein X^(c) isunsubstituted C₁-C₄ alkyl, halide, CCl₃, —OH, —SH, —CN, —OCH₃, or —CF₃.In embodiments, the compound is not a compound of formula (VIc) whereinX^(c) is R⁵. In embodiments, the compound is not

wherein X^(c) and X^(d) are independently a halide. In embodiments, thecompound is not a compound of formula (VId) wherein X^(c) and X^(d) areindependently a halide or —CH₃. In embodiments, the compound is not acompound of formula (VId) wherein X^(c) and X^(d) are independently ahalide, —CH₃, or —CF₃. In embodiments, the compound is not a compound offormula (VId) wherein X^(c) and X^(d) are independently a halide, —CH₃,—CCl₃, or —CF₃. In embodiments, the compound is not a compound offormula (VId) wherein X^(c) and X^(d) are independently a halide, —CH₃,—CCl₃, —CN, or —CF₃. In embodiments, the compound is not a compound offormula (VId) wherein X^(c) and X^(d) are independently a halide, —CH₃,—CCl₃, —OH, —CN, or —CF₃. In embodiments, the compound is not a compoundof formula (VId) wherein X^(c) and X^(d) are independently a halide,—CH₃, —CCl₃, —OH, —SH, —CN, —OCH₃, or —CF₃. In embodiments, the compoundis not a compound of formula (VId) wherein X^(c) and X^(d) areindependently a unsubstituted C₁-C₂ alkyl, halide, —CCl₃, —OH, —SH, —CN,—OCH₃, or —CF₃. In embodiments, the compound is not a compound offormula (VId) wherein X^(c) and X^(d) are independently a unsubstitutedC₁-C₃ alkyl, halide, —CCl₃, —OH, —SH, —CN, —OCH₃, or —CF₃. Inembodiments, the compound is not a compound of formula (VId) whereinX^(c) and X^(d) are independently a unsubstituted C₁-C₄ alkyl, halide,—CCl₃, —OH, —SH, —CN, —OCH₃, or —CF₃. In embodiments, the compound isnot a compound of formula (VId) wherein X^(c) and X^(d) areindependently an R⁵.

In embodiments, the compound is not

In embodiments, the compound is not

wherein A^(e) is a halide. In embodiments, the compound is not

wherein X^(e) is a halide.

In embodiments, the compound is not

wherein X^(e) is a halide and L² and L⁴ are a bond or unsubstitutedC₁-C₂ alkylene. In embodiments, the compound is not a compound offormula (VIId) wherein X^(e) is a halide and L² and L⁴ are a bond orunsubstituted C₁-C₃ alkylene. In embodiments, the compound is not acompound of formula (VIId) wherein X^(e) is a halide and L² and L⁴ are abond or unsubstituted C₁-C₄ alkylene.

In embodiments, the compound is not

wherein X^(e) is a halide and L² and L⁴ are a bond or unsubstitutedC₁-C₂ alkylene and L¹ and L³ are a bond or unsubstituted C₁-C₂ alkylene.In embodiments, the compound is not a compound of formula (VIIe) whereinX^(e) is a halide and L² and L⁴ are a bond or unsubstituted C₁-C₂alkylene and L¹ and L³ are a bond or unsubstituted C₁-C₃ alkylene. Inembodiments, the compound is not a compound of formula (VIIe) whereinX^(e) is a halide and L² and L⁴ are a bond or unsubstituted C₁-C₂alkylene and L¹ and L³ are a bond or unsubstituted C₁-C₄ alkylene. Inembodiments, the compound is not a compound of formula (VIIe) whereinX^(e) is a halide and L² and L⁴ are a bond or unsubstituted C₁-C₃alkylene and L¹ and L³ are a bond or unsubstituted C₁-C₃ alkylene. Inembodiments, the compound is not a compound of formula (VIIe) whereinX^(e) is a halide and L² and L⁴ are a bond or unsubstituted C₁-C₄alkylene and L¹ and L³ are a bond or unsubstituted C₁-C₄ alkylene. Inembodiments, the compound is not a compound of formula VIIa, VIIb, VIIc,VIId, or VIIe wherein X^(e) is an unsubstituted methyl or halide. Inembodiments, the compound is not a compound of formula VIIa, VIIb, VIIc,VIId, or VIIe wherein X^(e) is a unsubstituted C₁-C₂ alkyl, halide, or—CF₃. In embodiments, the compound is not a compound of formula VIIa,VIIb, VIIc, VIId, or VIIe wherein X^(e) is a unsubstituted C₁-C₄ alkyl,halide, —CCl₃, or —CF₃. In embodiments, the compound is not a compoundof formula VIIa, VIIb, VIIc, VIId, or VIIe wherein X^(e) is aunsubstituted C₁-C₄ alkyl, halide, —CCl₃, —CN, or —CF₃. In embodiments,the compound is not a compound of formula VIIa, VIIb, VIIc, VIId, orVIIe wherein X^(e) is a unsubstituted C₁-C₄ alkyl, halide, CCl₃, —OH,—SH, —CN, —OCH₃, or —CF₃. In embodiments, the compound is not a compoundof formula VIIa, VIIb, VIIc, VIId, or VIIe wherein X^(e) is an R⁵.

In embodiments, the compound is not

wherein L^(2B) is substituted or unsubstituted C₁-C₄ alkylene; L^(4B) issubstituted or unsubstituted C₁-C₄ alkylene; X^(f) is halide, C₁-C₄substituted or unsubstituted alkyl, C₁-C₄ or substituted orunsubstituted alkoxy; X^(g) is hydrogen, substituted or unsubstitutedC₁-C₄ alkyl, C₁-C₄ or substituted or unsubstituted alkoxy. Inembodiments, L^(2B) is substituted or unsubstituted C₁-C₃ alkylene. Inembodiments, L^(2B) is substituted or unsubstituted C₁-C₂ alkylene. Inembodiments, L^(2B) is substituted or unsubstituted methylene. Inembodiments, L^(2B) is unsubstituted methylene. In embodiments, L^(2B)is methylene substituted with unsubstituted C₁-C₄ alkyl. In embodiments,L^(2B) is methylene substituted with unsubstituted C₁-C₃ alkyl. Inembodiments, L^(2B) is methylene substituted with unsubstituted C₁-C₂alkyl. In embodiments, L^(2B) is methylene substituted withunsubstituted methyl. In embodiments, L^(2B) is methylene substitutedwith one unsubstituted methyl. In embodiments, L^(4B) is substituted orunsubstituted C₁-C₃ alkylene. In embodiments, L^(4B) is substituted orunsubstituted C₁-C₂ alkylene. In embodiments, L^(4B) is substituted orunsubstituted methylene. In embodiments, L^(4B) is unsubstitutedmethylene. In embodiments, L^(4B) is methylene substituted withunsubstituted C₁-C₄ alkyl. In embodiments, L^(4B) is methylenesubstituted with unsubstituted C₁-C₃ alkyl. In embodiments, L^(4B) ismethylene substituted with unsubstituted C₁-C₂ alkyl. In embodiments,L^(4B) is methylene substituted with unsubstituted methyl. Inembodiments, L^(4B) is methylene substituted with one unsubstitutedmethyl. In embodiments, X^(f) is halide. In embodiments, X^(f) is —Cl.In embodiments, X^(f) is —F. In embodiments, X^(f) is —Br. Inembodiments, X^(f) is —I. In embodiments, X^(f) is substituted orunsubstituted C₁-C₄ alkyl. In embodiments, X^(f) is substituted orunsubstituted C₁-C₃ alkyl. In embodiments, X^(f) is substituted orunsubstituted C₁-C₂ alkyl. In embodiments, X^(f) is substituted orunsubstituted methyl. In embodiments, X^(f) is unsubstituted methyl. Inembodiments, X^(f) is unsubstituted C₁-C₄ alkyl. In embodiments, X^(f)is unsubstituted C₁-C₃ alkyl. In embodiments, X^(f) is unsubstitutedC₁-C₂ alkyl. In embodiments, X^(f) is substituted or unsubstituted C₁-C₄alkoxy. In embodiments, X^(f) is substituted or unsubstituted C₁-C₃alkoxy. In embodiments, X^(f) is substituted or unsubstituted C₁-C₂alkoxy. In embodiments, X^(f) is substituted or unsubstituted methoxy.In embodiments, X^(f) is unsubstituted methoxy. In embodiments, X^(f) isunsubstituted C₁-C₄ alkoxy. In embodiments, X^(f) is unsubstituted C₁-C₃alkoxy. In embodiments, X^(f) is unsubstituted C₁-C₂ alkoxy. Inembodiments, X^(g) is halide. In embodiments, X^(g) is —Cl. Inembodiments, X^(g) is —F. In embodiments, X^(g) is —Br. In embodiments,X^(g) is —I. In embodiments, X^(g) is substituted or unsubstituted C₁-C₄alkyl. In embodiments, X^(g) is substituted or unsubstituted C₁-C₃alkyl. In embodiments, X^(g) is substituted or unsubstituted C₁-C₂alkyl. In embodiments, X^(g) is substituted or unsubstituted methyl. Inembodiments, X^(g) is unsubstituted methyl. In embodiments, X^(g) isunsubstituted C₁-C₄ alkyl. In embodiments, X^(g) is unsubstituted C₁-C₃alkyl. In embodiments, X^(g) is unsubstituted C₁-C₂ alkyl. Inembodiments, X^(g) is substituted or unsubstituted C₁-C₄ alkoxy. Inembodiments, X^(g) is substituted or unsubstituted C₁-C₃ alkoxy. Inembodiments, X^(g) is substituted or unsubstituted C₁-C₂ alkoxy. Inembodiments, X^(g) is substituted or unsubstituted methoxy. Inembodiments, X^(g) is unsubstituted methoxy. In embodiments, X^(g) isunsubstituted C₁-C₄ alkoxy. In embodiments, X^(g) is unsubstituted C₁-C₃alkoxy. In embodiments, X^(g) is unsubstituted C₁-C₂ alkoxy. Inembodiments, X^(g) is hydrogen. In embodiments, the compound is not

In embodiments, the compound is not

wherein X^(g) is H, —Cl, —CH₃, or —OCH₃. In embodiments, the compound isnot

In embodiments, the compound is not a compound selected from the groupconsisting of:

In embodiments, the compound is not a compound selected from the groupconsisting of the compounds of Table 2.

In embodiments, the compound is an inhibitor of the integrated stressresponse. In embodiments, the compound is an inhibitor of a pathwayactivated by eIF2α phosphorylation. In embodiments, the compound is aninhibitor of a pathway activated by PERK activity. In embodiments, thecompound is an inhibitor of a pathway activated by accumulation ofunfolded protins in the endoplasmic reticulum. In embodiments, thecompound is an inhibitor of a pathway activated by GCN2 activity. Inembodiments, the compound is an inhibitor of a pathway activated byamino acid starvation. In embodiments, the compound is an inhibitor of apathway activated by PKR activity. In embodiments, the compound is aninhibitor of a pathway activated by viral infection. In embodiments, thecompound is an inhibitor of a pathway activated by HRI activity. Inembodiments, the compound is an inhibitor of a pathway activated by hemedeficiency. In embodiments, the compound is an inhibitor of a pathwaythat decreases bulk protein synthesis and includes eIF2α. Inembodiments, the compound is an inhibitor of a pathway activated byATF4. In embodiments, the compound is an inhibitor of a pathwayactivated by CHOP activity. In embodiments, the compound is an activatorof apoptosis. In embodiments, the compound increases the level ofapoptosis relative to the level of apoptosis in the absence of thecompound. In embodiments, the compound is an inhibitor of a pathwayactivated by hypoxic conditions that includes eIF2α. In embodiments, thecompound is an inhibitor of a pathway downstream of eIF2αphosphorylation. In embodiments, the compound is an inhibitor of apathway downstream of eIF2α phosphorylation of serine 51 (in the humanprotein or the corresponding residue in a non-human protein). Inembodiments, the compound is an inhibitor of a pathway downstream ofeIF2α phosphorylation by PERK, GCN2, PKR, or HRI. In embodiments, thecompound is an inhibitor of neuronal cell death. In embodiments, thecompound is a cytotoxic agent. In embodiments, the compound is ananti-cancer agent. In embodiments, the compound is an inhibitor of aprotein activated by eIF2α phosphorylation (directly or indirectly). Inembodiments, the compound is an inhibitor of a protein, wherein thelevel of protein (e.g. amount or activity level) is increased by eIF2αphosphorylation (directly or indirectly). In embodiments, the compoundincreases caspase 3 activity. In embodiments, the compound increasescaspase 7 activity. In embodiments, the compound increases apoptosis incells under ER stress. In embodiments, the compound increases apoptosisin cells under ER stress but not cells under the same conditions exceptthat they are not under ER stress. In embodiments, the compoundincreases apoptosis in cells under ER stress more than in cells underthe same conditions except that they are not under ER stress. Inembodiments, the compound inhibits the formation of the eIF2 complex.

In embodiments, R¹ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R^(H)-substituted or unsubstituted alkyl, R¹¹-substituted orunsubstituted heteroalkyl, R¹¹-substituted or unsubstituted cycloalkyl,R¹¹-substituted or unsubstituted heterocycloalkyl, R¹¹-substituted orunsubstituted aryl, or R¹¹-substituted or unsubstituted heteroaryl.

In embodiments, R³ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R^(H)-substituted or unsubstituted alkyl, R¹³-substituted orunsubstituted heteroalkyl, R¹³-substituted or unsubstituted cycloalkyl,R¹³-substituted or unsubstituted heterocycloalkyl, R¹³-substituted orunsubstituted aryl, or R¹³-substituted or unsubstituted heteroaryl.

In embodiments, R⁵ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁵-substituted or unsubstituted alkyl, R¹⁵-substituted or unsubstitutedheteroalkyl, R¹⁵-substituted or unsubstituted cycloalkyl,R¹⁵-substituted or unsubstituted heterocycloalkyl, R¹⁵-substituted orunsubstituted aryl, or R¹⁵-substituted or unsubstituted heteroaryl.

In embodiments, R⁶ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁶-substituted or unsubstituted alkyl, R¹⁶-substituted or unsubstitutedheteroalkyl, R¹⁶-substituted or unsubstituted cycloalkyl,R¹⁶-substituted or unsubstituted heterocycloalkyl, R¹⁶-substituted orunsubstituted aryl, or R¹⁶-substituted or unsubstituted heteroaryl.

In embodiments, R⁷ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —S O₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R′⁷-substituted or unsubstituted alkyl, R¹⁷-substituted or unsubstitutedheteroalkyl, R¹⁷-substituted or unsubstituted cycloalkyl,R¹⁷-substituted or unsubstituted heterocycloalkyl, R¹⁷-substituted orunsubstituted aryl, or R¹⁷-substituted or unsubstituted heteroaryl.

In embodiments, R⁸ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁸-substituted or unsubstituted alkyl, R¹⁸-substituted or unsubstitutedheteroalkyl, R¹⁸-substituted or unsubstituted cycloalkyl,R¹⁸-substituted or unsubstituted heterocycloalkyl, R¹⁸-substituted orunsubstituted aryl, or R¹⁸-substituted or unsubstituted heteroaryl.

In embodiments, R⁹ is independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,R¹⁹-substituted or unsubstituted alkyl, R¹⁹-substituted or unsubstitutedheteroalkyl, R¹⁹-substituted or unsubstituted cycloalkyl,R¹⁹-substituted or unsubstituted heterocycloalkyl, R¹⁹-substituted orunsubstituted aryl, or R¹⁹-substituted or unsubstituted heteroaryl.

In embodiments, L¹ is independently a bond, R²¹-substituted orunsubstituted alkylene, or R²¹-substituted or unsubstitutedheteroalkylene.

In embodiments, L² is independently a bond, R²²-substituted orunsubstituted alkylene, or R²²-substituted or unsubstitutedheteroalkylene.

In embodiments, L^(2B) is independently a bond or R^(22B)-substituted orunsubstituted alkylene.

In embodiments, L³ is independently a bond, R²³-substituted orunsubstituted alkylene, or R²³-substituted or unsubstitutedheteroalkylene.

In embodiments, L⁴ is independently a bond, R²⁴-substituted orunsubstituted alkylene, or R²⁴-substituted or unsubstitutedheteroalkylene.

In embodiments, L^(4B) is independently a bond or R^(24B)-substituted orunsubstituted alkylene.

In embodiments, ring A is independently an R²⁵-substituted orunsubstituted cycloalkylene, or R²⁵-substituted or unsubstitutedarylene.

Each R¹¹, R¹³, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²¹, R²², R^(22B), R²³, R²⁴,R^(24B), and R²⁵ is independently hydrogen, oxo,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH, —CH₂CCH,—SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, —OCH₂CCH, —NHC(O)CH₃, —NHCH₃, —NHC(S)CH₃, —N(CH₃)₂, —C(O)NHNH₂,unsubstituted alkyl, unsubstituted heteroalkyl, unsubstitutedcycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, orunsubstituted heteroaryl. In embodiments, R²² is —CF₃. In embodiments,R^(22B) is —CF₃. In embodiments, R²⁴ is —CF₃. In embodiments, R^(24B) is—CF₃.

In some embodiments, a compound as described herein may include multipleinstances of R⁵, R⁶, R⁷, R, R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷,R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R^(22B), R^(24B) and/or othervariables. In such embodiments, each variable may optional be differentand be appropriately labeled to distinguish each group for greaterclarity. For example, where each R⁵, R⁶, R⁷, R, R⁹, R¹⁰, R¹¹, R¹², R¹³,R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R^(22B),and/or R^(24B), is different, they may be referred to, for example, asR^(5.1), R^(5.2), R^(5.3), R^(5.4), R^(5.5), R^(6.1), R^(6.2), R^(6.3),R^(6.4), R^(6.5), R^(7.1), R^(7.2), R^(7.3), R^(7.4), R^(7.5), R^(8.1),R^(8.2), R^(8.3), R^(8.4), R^(8.5), R^(9.1), R^(9.2), R^(9.3), R^(9.4),R^(9.5), R^(10.1), R^(10.2), R^(10.3), R^(10.4), R^(10.5), R^(11.1),R^(11.3), R^(11.4), R^(11.5), R^(12.1), R^(12.2), R^(12.3), R^(12.4),R^(12.5), R^(13.1), R^(13.2), R^(13.3), R^(13.4), R^(13.5), R^(14.1),R^(14.3), R^(14.4), R^(14.5), R^(15.1), R^(15.2), R^(15.3), R^(15.4),R^(15.5), R^(16.1), R^(16.2), R^(16.3), R^(16.4), R^(16.5), R^(17.1),R^(17.2), R^(17.3), R^(17.4), R^(17.5), R^(18.1), R^(18.2), R^(18.3),R^(18.4), R^(18.5), R^(19.1), R^(19.2), R^(19.3), R^(19.4), R^(19.5),R^(2.1), R^(2.2), R^(2.3), R^(2.4), R^(2.5), R^(21.1), R^(21.2),R^(21.3), R^(21.4), R^(21.5), R^(22.1), R^(22.2), R^(22.3), R^(22.4),R^(22.5), R^(23.1), R^(23.2), R^(23.3), R^(23.4), R^(23.5), R^(24.1),R^(24.2), R^(24.3), R^(24.4), R^(24.5), R^(25.1), R^(25.2), R^(25.3),R^(25.4), R^(25.5), R^(22B.1), R^(22B.2), R^(22B.3), R^(22B.4),R^(22B.5), R^(24B1), R^(24B.2), R^(24B.3), R^(24B.4), and/or R^(24B.5),respectively, wherein the definition of R⁵ is assumed by R^(5.1),R^(5.2), R^(5.3), R^(5.4), and/or R^(5.5), the definition of R⁶ isassumed by R^(6.1), R^(6.2), R^(6.3), R^(6.4), and/or R^(6.5), thedefinition of R⁷ is assumed by R^(7.1), R^(7.2), R^(7.3), R^(7.4),and/or R^(7.5), the definition of R⁸ is assumed by R^(8.1), R^(8.2),R^(8.3), R^(8.4), and/or R^(8.5), the definition of R⁹ is assumed byR^(9.1), R^(9.2), R^(9.3), R^(9.4), and/or R^(9.5), the definition ofR¹⁰ is assumed by R^(10.1), R^(10.2), R^(10.3), R^(10.4), and/orR^(10.5), the definition of R¹¹ is assumed by R^(11.1), R^(11.2),R^(11.3), R^(11.4), and/or R^(11.5), the definition of R¹² is assumed byR^(12.1), R^(12.2), R^(12.3), R^(12.4), and/or R^(12.5), the definitionof R¹³ is assumed by R^(13.1), R^(13.2), R^(13.3), R^(13.4), and/orR^(13.5), the definition of R¹⁴ is assumed by R^(14.1), R^(14.2),R^(14.3), R^(14.4), and/or R^(14.5), the definition of R¹⁵ is assumed byR^(15.1), R^(15.2), R^(15.3), R^(15.4), and/or R^(15.5), the definitionof R¹⁶ is assumed by R^(16.1), R^(16.2), R^(16.3), R^(16.4), and/orR^(16.5), the definition of R¹⁷ is assumed by R^(17.1), R^(17.2),R^(17.3), R^(17.4), and/or R^(17.5), the definition of R¹⁸ is assumed byR^(18.1), R^(18.2), R^(18.3), R^(18.4), and/or R^(18.5), the definitionof R¹⁹ is assumed by R^(19.1), R^(19.2), R^(19.3), R^(19.4), and/orR^(19.5), the definition of R²⁰ is assumed by R^(20.1), R^(20.2),R^(20.3), R^(20.4), and/or R^(20.5), the definition of R²¹ is assumed byR^(21.1), R^(21.2), R^(21.3), R^(21.4), and/or R^(21.5), the definitionof R²² is assumed by R^(22.1), R^(22.2), R^(22.3), R^(22.4), and/orR^(22.5), the definition of R²³ is assumed by R^(23.1), R^(23.2),R^(23.3), R^(23.4), and/or R^(23.5), the definition of R²⁴ is assumed byR^(24.1), R^(24.2), R^(24.3), R^(24.4), and/or R^(24.5), the definitionof R²⁵ is assumed by R^(25.1), R^(25.2), R^(25.3), R^(25.4), and/orR^(25.5), the definition of R^(22B) is assumed by R^(22B.1), R^(22B.2),R^(22B.3), R^(22B.4), and/or R^(22B.5), and the definition of R^(24B) isassumed by R^(24B.1), R^(24B.2), R^(24B.3), R^(24B.4), and/or R^(24B.5).The variables used within a definition of R⁵, R⁶, R⁷, R⁸, R⁹, R¹, R¹¹,R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R², R²¹, R²², R²³, R²⁴, R²⁵,R^(22B), R^(24B), and/or other variables that appear at multipleinstances and are different may similarly be appropriately labeled todistinguish each group for greater clarity.

In embodiments, the compounds set forth herein are provided aspharmaceutical compositions including the compound and apharmaceutically acceptable excipient.

In some embodiments, the compound is a compound described herein (e.g.compound of formula I, Ia, II, III, IIIa, IIIb, or IV, or any embodimentthereof, including compounds described for use in a method). In someembodiments, the compound is a compound described in the Examples, anexample, a table, the figures, or a figure. In some embodiments, thecompound is a compound described in Table 2.

Pharmaceutical Compositions

In another aspect is provided a pharmaceutical composition including apharmaceutically acceptable excipient and a compound, orpharmaceutically acceptable salt thereof, as described herein, includingembodiments (e.g. compound of formula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih,II, III, IIIa, IIIb, IIIc, or IV, or any embodiment thereof, includingcompounds described for use in a method herein or in the Compoundssection above or in an example, table, figure, or claim). In someembodiments, the compound is a compound described in Table 2. Inembodiments of the pharmaceutical compositions, the compound, orpharmaceutically acceptable salt thereof, as described herein, includingembodiments (e.g. compound of formula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih,II, III, IIIa, IIIb, IIIc, or IV, or any embodiment thereof, includingcompounds described for use in a method herein or in the Compoundssection above or in an example, table, figure, or claim) is included ina therapeutically effective amount.

In embodiments of the pharmaceutical compositions, the pharmaceuticalcomposition includes a second agent (e.g. therapeutic agent). Inembodiments of the pharmaceutical compositions, the pharmaceuticalcomposition includes a second agent (e.g. therapeutic agent) in atherapeutically effective amount. In embodiments of the pharmaceuticalcompositions, the second agent is an agent for treating cancer (e.g.pancreatic cancer, breast cancer, multiple myeloma, or cancers ofsecretory cells), neurodegenerative diseases, vanishing white matterdisease, childhood ataxia with CNS hypo-myelination, and/or intellectualdisability syndromes (e.g. associated with impaired function of eIF2 orcomponents in a signal transduction pathway including eIF2). Inembodiments, the second agent is an anti-cancer agent. In embodiments,the second agent is a chemotherapeutic. In embodiments, the second agentis an agent for improving memory. In embodiments, the second agent is anagent for treating a neurodegenerative disease. In embodiments, thesecond agent is an agent for treating vanishing white matter disease. Inembodiments, the second agent is an agent for treating childhood ataxiawith CNS hypo-myelination. In embodiments, the second agent is an agentfor treating an intellectual disability syndrome. In embodiments, thesecond agent is an agent for treating pancreatic cancer. In embodiments,the second agent is an agent for treating breast cancer. In embodiments,the second agent is an agent for treating multiple myeloma. Inembodiments, the second agent is an agent for treating myeloma. Inembodiments, the second agent is an agent for treating a cancer of asecretory cell. In embodiments, the second agent is an agent forreducing eIF2α phosphorylation. In embodiments, the second agent is anagent for inhibiting a pathway activated by eIF2α phosphorylation. Inembodiments, the second agent is an agent for inhibiting a pathwayactivated by eIF2α. In embodiments, the second agent is an agent forinhibiting the integrated stress response. In embodiments, the secondagent is an anti-inflammatory agent. In embodiments, the second agent isan agent for treating postsurgical cognitive dysfunction (POCD). Inembodiments, the second agent is an agent for treating traumatic braininjury (TBI).

Additional Embodiments

1p. A method of treating a disease in a patient in need of suchtreatment, said method comprising administering a therapeuticallyeffective amount of a compound, or a pharmaceutically acceptable saltthereof, to said patient, wherein said disease is selected from thegroup consisting of cancer, a neurodegenerative disease, vanishing whitematter disease, childhood ataxia with CNS hypo-myelination, and anintellectual disability syndrome; and wherein said compound has theformula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ areindependently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR′, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

2p. The method of embodiment 1p, wherein the compound has the formula:

3p. The method of any one of embodiments 1p to 2p, wherein L¹ and L³ areindependently a bond or substituted or unsubstituted alkylene.

4p. The method of any one of embodiments 1p to 3p, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₅ alkylene.

5p. The method of any one of embodiments 1p to 4p, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₃ alkylene.

6p. The method of any one of embodiments 1p to 5p, wherein L¹ and L³ areindependently substituted or unsubstituted methylene.

7p. The method of any one of embodiments 1p to 3p, wherein L¹ and L³ areindependently a bond.

8p. The method of any one of embodiments 1p to 3p, wherein L¹ and L³ areindependently an unsubstituted alkylene.

9p. The method of embodiment 1p, wherein the compound has the formula:

wherein, R⁸ and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; and b and d are independently 0or 1.

10p. The method of embodiment 9p, wherein the compound has the formula:

11p. The method of any one of embodiments 9p to 10p, wherein b and d are1.

12p. The method of any one of embodiments 9p to 11p, wherein R⁸ and R⁹are hydrogen.

13p. The method of embodiment 1p, wherein the compound has the formula:

14p. The method of any one of embodiments 1p to 13p, wherein R¹ and R³are hydrogen.

15p. The method of any one of embodiments 1p to 14p, wherein R² and R⁴are ═0.

16p. The method of any one of embodiments 1p to 15p, wherein L² isL^(2A)-L^(2B)-L^(2C), wherein L^(2A) is bonded to the substituted orunsubstituted phenyl; L^(2A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—; L^(2B) is a bond or substituted or unsubstituted alkylene; andL^(2C) is a bond, —O—, or —NH—.

17p. The method of embodiment 16p, wherein L^(2A) is bonded to thesubstituted or unsubstituted phenyl; L^(2A) is a bond; L^(2B) isunsubstituted methylene; and L^(2C) is —O—.

18p. The method of any one of embodiments 1p to 17p, wherein L⁴ isL^(4A)-L^(4B)-L^(4C) wherein L^(4A) is bonded to the substituted orunsubstituted phenyl; L^(4A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—; L^(4B) is a bond or substituted or unsubstituted alkylene; andL^(4c) is a bond, —O—, or —NH—.

19p. The method of embodiment 18p, wherein L^(4A) is bonded to thesubstituted or unsubstituted phenyl; L^(4A) is a bond; L^(4B) isunsubstituted methylene; and L^(4C) is —O—.

20p. The method of any one of embodiments 1p to 19p, wherein R⁵ and R⁶are independently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

21p. The method of any one of embodiments 1p to 20p, wherein z5 and z6are independently 0 to 2.

22p. The method of any one of embodiments 1p to 21p, wherein thecompound is

23p. The method of any one of embodiments 1p to 22p, wherein the diseaseis cancer.

24p. The method of any one of embodiments 1p to 22p, wherein the diseaseis a neurodegenerative disease.

25p. The method of any one of embodiments 1p to 22p, wherein the diseaseis vanishing white matter disease.

26p. The method of any one of embodiments 1p to 22p, wherein the diseaseis childhood ataxia with CNS hypo-myelination.

2′7p. The method of any one of embodiments 1p to 22p, wherein thedisease is associated with eIF2α phosphorylation.

28p. A method of increasing protein expression by a cell or in vitroexpression system, said method comprising administering an effectiveamount of a compound, or a pharmaceutically acceptable salt thereof, tosaid cell or expression system, wherein said compound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ areindependently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

29p. The method of embodiment 28p, wherein the compound has the formula:

30p. A method of improving long-term memory in a patient, said methodcomprising administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, to saidpatient, wherein said compound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ areindependentlyhydrogen; halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN,—S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂,—CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

31p. The method of embodiment 30p, wherein said compound has theformula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ areindependently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; and z5 and z6 are independently an integer from 0 to 5.

32p. A compound, or a pharmaceutically acceptable salt thereof, havingthe formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, substituted or unsubstituted alkylene orsubstituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ areindependently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO 4H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5; with the proviso that the compoundis not

33p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 32p, wherein said compound has the formula:

34p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 33p, wherein L¹ and L³ are independently abond or substituted or unsubstituted alkylene.

35p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 34p, wherein L¹ and L³ are independentlysubstituted or unsubstituted C₁-C₅ alkylene.

36p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 35p, wherein L¹ and L³ are independentlysubstituted or unsubstituted C₁-C₃ alkylene.

3′7p. The compound, or a pharmaceutically acceptable salt thereof, ofany one of embodiments 32p to 36p, wherein L¹ and L³ are independentlysubstituted or unsubstituted methylene.

38p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 34p, wherein L¹ and L³ are independently abond.

39p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 34p, wherein L′ and L³ are independently anunsubstituted alkylene.

40p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 32p, wherein the compound has the formula:

wherein, R⁸ and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; and b and d are independently 0or 1.

41p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 40p, wherein the compound has the formula:

42p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 40p to 41p, wherein b and d are 1.

43p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 40p to 42p, wherein R⁸ and R⁹ are hydrogen.

44p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 32p, wherein the compound has the formula:

45p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 44p, wherein R¹ and R³ are hydrogen.

46p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 45p, wherein R² and R⁴ are ═O.

47p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 46p, wherein L² is L^(2A)-L^(2B)-L^(2C),wherein L^(2A) is bonded to the substituted or unsubstituted phenyl;L^(2A) is a bond, —O—, —S—, —NH—, —S(O)—, or —S(O)₂—; L^(2B) is a bondor substituted or unsubstituted alkylene; and L^(2C) is a bond, —O—, or—NH—.

48p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 47p, wherein L^(2A) is bonded to the substituted orunsubstituted phenyl; L^(2A) is a bond; L^(2B) is unsubstitutedmethylene; and L^(2C) is —O—.

49p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 48p, wherein L⁴ is L^(4A)-L^(4B)-L^(4C), Lwherein L^(4A) is bonded to the substituted or unsubstituted phenyl;L^(4A) is a bond, —O—, —S—, —NH—, —S(O)—, or —S(O)₂—; L^(4B) is a bondor substituted or unsubstituted alkylene; and L^(4C) is a bond, —O—, or—NH—.

50p. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 49p, wherein L^(4A) is bonded to the substituted orunsubstituted phenyl; L^(4A) is a bond; L^(4B) is unsubstitutedmethylene; and L^(4C) is —O—.

51p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 48p, wherein R⁵ and R⁶ are independentlyhydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

52p. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 32p to 51p, wherein z5 and z6 are independently 0 to2.

53p. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound, or pharmaceutically acceptable saltthereof, of any one of embodiments 32p to 52p.

1. A compound, or a pharmaceutically acceptable salt thereof, having theformula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5; with the proviso that the compoundis not

2. The compound of embodiment 1, wherein said compound has the formula:

3. The compound of any one of embodiments 1 to 2, wherein L¹ and L³ areindependently a bond or substituted or unsubstituted alkylene.

4. The compound of any one of embodiments 1 to 3, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₅ alkylene.

5. The compound of any one of embodiments 1 to 4, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₃ alkylene.

6. The compound of any one of embodiments 1 to 5, wherein L¹ and L³ areindependently substituted or unsubstituted methylene.

7. The compound of any one of embodiments 1 to 3, wherein L¹ and L³ areindependently a bond.

8. The compound of any one of embodiments 1 to 3, wherein L¹ and L³ areindependently an unsubstituted alkylene.

9. The compound of embodiment 1, wherein the compound has the formula:

wherein, R⁸ and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; and b and d are independently 0or 1.

10. The compound of embodiment 9, wherein the compound has the formula:

11. The compound of any one of embodiments 9 to 10, wherein b and d are1.

12. The compound of any one of embodiments 9 to 11, wherein R⁸ and R⁹are hydrogen.

13. The compound of one of embodiments 1 to 8, wherein the compound hasthe formula:

14. The compound of any one of embodiments 1 to 13, wherein R¹ and R³are hydrogen.

15. The compound of any one of embodiments 1 to 14, wherein R² and R⁴are ═0.

16. The compound of any one of embodiments 1 to 15, wherein L² isL^(2A)-L^(2B)-L^(2C), wherein L^(2A) is bonded to the substituted orunsubstituted phenyl; L^(2A) is a bond, —CO—, —S—, —NH—, —S(O)—, or—S(O)₂—; L^(2B) is a bond or substituted or unsubstituted alkylene; andL² is a bond, —O—, or —NH—.

17. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 16, wherein L^(2A) is bonded to the substituted orunsubstituted phenyl; L^(2A) is a bond; L^(2B) is unsubstitutedmethylene; and L^(2C) is —O—.

18. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 1 to 17, wherein L⁴ is L^(4A)-L^(4B)-L^(4C), whereinL^(4A) is bonded to the substituted or unsubstituted phenyl; L^(4A) is abond, —O—, —S—, —NH—, —S(O)—, or —S(O)₂—; L^(4B) is a bond orsubstituted or unsubstituted alkylene; and L^(4C) is a bond, —O—, or—NH—.

19. The compound, or a pharmaceutically acceptable salt thereof, ofembodiment 18, wherein L^(4A) is bonded to the substituted orunsubstituted phenyl; L^(4A) is a bond; L^(4B) is unsubstitutedmethylene; and L^(4C) is —O—.

20. The compound, or a pharmaceutically acceptable salt thereof, of anyone of embodiments 1 to 19, wherein R⁵ and R⁶ are independentlyhydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

21. The compound of any one of embodiments 1 to 20, wherein z5 and z6are independently 0 to 2.

22. The compound of embodiment 21, wherein the compound has the formula:

R^(5.1) and R^(6.1) are independently hydrogen, halogen, —CF₃, —CN, —N₃,substituted or unsubstituted C₁-C₄ alkyl, substituted or unsubstituted 2to 4 membered heteroalkyl, substituted or unsubstituted 5 to 6 memberedheteroaryl,

R^(5.2) and R^(6.2) are independently hydrogen, halogen, —CCSi(CH₃)₃,—CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted orunsubstituted 5 to 6 membered heteroaryl,

23. The compound of embodiment 22, wherein, R^(5.1) and R^(6.1) areindependently halogen, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl; R^(5.2) and R^(6.2) are independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl.

24. The compound of embodiment 23, wherein, R^(5.1) and R^(6.1) areindependently —Cl, —I, —CF₃, —CH₃, or —CCH; R^(5.2) and R^(6.2) areindependently hydrogen, —Cl, —F, —I, —CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or—CCH.

25. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound, or pharmaceutically acceptable saltthereof, of any one of embodiments 1 to 24.

26. A method of treating an integrated stress response-associateddisease in a patient in need of such treatment, said method comprisingadministering a therapeutically effective amount of a compound, or apharmaceutically acceptable salt thereof, to said patient, wherein saidcompound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

27. A method of treating a disease associated with phosphorylation ofeIF2α in a patient in need of such treatment, said method comprisingadministering a therapeutically effective amount of a compound, or apharmaceutically acceptable salt thereof, to said patient, wherein saidcompound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

28. The method of one of embodiments 26 to 27, wherein said disease iscancer, a neurodegenerative disease, vanishing white matter disease,childhood ataxia with CNS hypo-myelination, or an intellectualdisability syndrome.

29. The method of embodiment 28, wherein the disease is cancer.

30. The method of embodiment 28, wherein the disease is aneurodegenerative disease.

31. The method of embodiment 28, wherein the disease is vanishing whitematter disease.

32. The method of embodiment 28, wherein the disease is childhood ataxiawith CNS hypo-myelination.

33. The method of embodiment 28, wherein the disease is an intellectualdisability syndrome.

34. A method of treating an inflammatory disease in a patient in need ofsuch treatment, said method comprising administering a therapeuticallyeffective amount of a compound, or a pharmaceutically acceptable saltthereof, to said patient, wherein said compound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —C ONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

35. The method of embodiment 34, wherein said inflammatory disease isassociated with neurological inflammation.

36. The method of one of embodiments 34 to 35, wherein said inflammatorydisease is postoperative cognitive dysfunction.

37. The method of one of embodiments 34 to 35, wherein said inflammatorydisease is traumatic brain injury.

38. A method of improving long-term memory in a patient, said methodcomprising administering a therapeutically effective amount of acompound, or a pharmaceutically acceptable salt thereof, to saidpatient, wherein said compound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH,—OCH₃, —OCF₃, —OCHF₂, —N₃, substituted or unsubstituted alkyl,substituted or unsubstituted heteroalkyl, substituted or unsubstitutedcycloalkyl, substituted or unsubstituted heterocycloalkyl, substitutedor unsubstituted aryl, or substituted or unsubstituted heteroaryl; R²and R⁴ are independently ═NR⁷, ═O, or ═S; z2 and z4 are independently 0or 1; and z5 and z6 are independently an integer from 0 to 5.

39. A method of increasing protein expression by a cell or in vitroexpression system, said method comprising administering an effectiveamount of a compound, or a pharmaceutically acceptable salt thereof, tosaid cell or expression system, wherein said compound has the formula:

wherein, ring A is substituted or unsubstituted cycloalkylene orsubstituted or unsubstituted arylene; L¹, L², L³, and L⁴ areindependently a bond, —NH—, —O—, —S—, —S(O)—, —S(O)₂—, substituted orunsubstituted alkylene or substituted or unsubstituted heteroalkylene;R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃,—C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂,—ONH₂, —NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCH₃, —OCF₃, —OCHF₂,—N₃, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; R² and R⁴ are independently═NR⁷, ═O, or ═S; z2 and z4 are independently 0 or 1; and z5 and z6 areindependently an integer from 0 to 5.

40. The method of one of embodiments 26 to 39, wherein the compound hasthe formula:

41. The method of any one of embodiments 26 to 40, wherein L¹ and L³ areindependently a bond or substituted or unsubstituted alkylene.

42. The method of any one of embodiments 26 to 41, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₅ alkylene.

43. The method of any one of embodiments 26 to 42, wherein L¹ and L³ areindependently substituted or unsubstituted C₁-C₃ alkylene.

44. The method of any one of embodiments 26 to 43, wherein L¹ and L³ areindependently substituted or unsubstituted methylene.

45. The method of any one of embodiments 26 to 41, wherein L¹ and L³ areindependently a bond.

46. The method of any one of embodiments 26 to 41, wherein L¹ and L³ areindependently an unsubstituted alkylene.

47. The method of one of embodiments 26 to 46, wherein the compound hasthe formula:

wherein, R⁸ and R⁹ are independently hydrogen,halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl; and b and d are independently 0or 1.

48. The method of embodiment 47, wherein the compound has the formula:

49. The method of any one of embodiments 47 to 48, wherein b and d are1.

50. The method of any one of embodiments 47 to 49, wherein R⁸ and R⁹ arehydrogen.

51. The method of one of embodiments 26 to 46, wherein the compound hasthe formula:

52. The method of any one of embodiments 26 to 51, wherein R¹ and R³ arehydrogen.

53. The method of any one of embodiments 26 to 51, wherein R² and R⁴ are═0.

54. The method of any one of embodiments 26 to 52, wherein L² isL^(2A)-L^(2B)-L^(2C), wherein L^(2A) is bonded to the substituted orunsubstituted phenyl; L^(2A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—; L^(2B) is a bond or substituted or unsubstituted alkylene; andL^(2C) is a bond, —O—, or —NH—.

55. The method of embodiment 54, wherein L^(2A) is bonded to thesubstituted or unsubstituted phenyl; L^(2A) is a bond; L^(2B) isunsubstituted methylene; and L^(2C) is —O—.

56. The method of any one of embodiments 26 to 55, wherein L⁴ isL^(4A)-L^(4B)-L^(4C) wherein L^(4A) is bonded to the substituted orunsubstituted phenyl; L^(4A) is a bond, —O—, —S—, —NH—, —S(O)—, or—S(O)₂—; L^(4B) is a bond or substituted or unsubstituted alkylene; andL^(4c) is a bond, —O—, or —NH—.

57. The method of embodiment 56, wherein L^(4A) is bonded to thesubstituted or unsubstituted phenyl; L^(4A) is a bond; L^(4B) isunsubstituted methylene; and L^(4C) is —O—.

58. The method of any one of embodiments 26 to 57, wherein R⁵ and R⁶ areindependently hydrogen,

halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH,—NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH,—CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂,—NHC═(O)NHNH₂,

—NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCH₃, —OCF₃,—OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstitutedheteroalkyl, substituted or unsubstituted cycloalkyl, substituted orunsubstituted heterocycloalkyl, substituted or unsubstituted aryl, orsubstituted or unsubstituted heteroaryl.

59. The method of any one of embodiments 26 to 58, wherein z5 and z6 areindependently 0 to 2.

60. The method of any one of embodiments 26 to 59, wherein the compoundhas the formula:

R^(5.1) and R^(6.1) are independently hydrogen, halogen, —CF₃, —CN, —N₃,substituted or unsubstituted C₁-C₄ alkyl, substituted or unsubstituted 2to 4 membered heteroalkyl, substituted or unsubstituted 5 to 6 memberedheteroaryl,

and R^(5.2) and R^(6.2) are independently hydrogen, halogen,—CCSi(CH₃)₃, —CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄alkyl, substituted or unsubstituted 2 to 4 membered heteroalkyl,substituted or unsubstituted 5 to 6 membered heteroaryl,

61. The method of embodiment 60, wherein, R^(5.1) and R^(6.1) areindependently halogen, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl; and R^(5.2) and R^(6.2) are independently hydrogen, halogen,—CCSi(CH₃)₃, —NO₂, unsubstituted C₁-C₃ alkyl, or unsubstituted C₁-C₃haloalkyl.

62. The method of embodiment 61, wherein, R^(5.1) and R^(6.1) areindependently —Cl, —I, —CF₃, —CH₃, or —CCH; and R^(5.2) and R^(6.2) areindependently hydrogen, —Cl, —F, —I, —CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or—CCH.

63. The method of any one of embodiments 26 to 62, wherein the compoundis

64. The method of any one of embodiments 26 to 62, wherein the compoundis

65. The compound of any one of embodiments 1 to 24, wherein the compoundis not

wherein X^(g) is H, —Cl, —CH₃, or —OCH₃; or

EXAMPLES

Phosphorylation of the α-subunit of initiation factor 2 (eIF2α) controlsprotein synthesis by a mechanism that is conserved in eukaryotic cells.In metazoa, four eIF2α kinases (PERK, PKR, GCN2, and HRI) are activatedby distinct stress conditions and converge on phosphorylating a uniqueserine in eIF2α. This collection of signaling pathways is termed“integrated stress response”, or ISR. eIF2α phosphorylation globallydiminishes protein synthesis but also allows a group of specializedmRNAs to become preferentially translated.

We identified novel small molecules (e.g. ISRIB) that render cellsresistant to the effects of eIF2α phosphorylation, restoring the cell'stranslation capacity. ISRIB is the first reported antagonist of the ISR.It acts as a potent and stereospecific inhibitor with an IC₅₀ of 5 nM incultured cells, suggesting a specific and tight interaction with itscellular target. By blocking signaling through the PERK branch of theUPR, ISRIB prevents cells from re-establishing ER homeostasis.Unmitigated ER stress synergizes with ISRIB to induce apoptosis.

ISM shows good pharmacokinetic properties and no overt toxicity in mice,making it suitable for in vivo studies. As such, ISRIB emerges as apowerful tool to explore the roles of the UPR and the ISR in diseasemodels and physiological processes. In particular, we utilized ISRIB toshow that overriding the consequences of eIF2α phosphorylation enhancesmemory consolidation in rodents, suggesting an important role of eIF2αphosphorylation in modulating higher-order brain function.

A. Compound Identification and Characterization

Starting with a cell-based, high-throughput screen for small moleculeinhibitors of PERK signaling, we identified a compound, named ISRIB,which potently (IC₅₀=5 nM) reverses the effects of eIF2αphosphorylation, effectively blunting its functional consequences.

Design of cell-based screen for inhibitors of PERK signaling. Byinterrogating a large chemical library for small molecules that blockPERK signaling, we identified ISRIB as a potent ISR inhibitor,functioning downstream of all eIF2α kinases. ISRIB proves a powerfultool to explore the consequences of acute inhibition of the ISR in cellsand animals.

To identify inhibitors of PERK signaling, we engineered a reporter thatallows monitoring of PERK activation in living cells. To this end, weconstructed a retroviral vector containing the open-reading frame offirefly luciferase fused to the 5′UTR of ATF4 mRNA (FIG. 1A), whichcontains two short open-reading frames (uORFs) that control ATF4translation in a stress-dependent manner. After infection, weestablished a HEK293T cell line harboring the stably integratedreporter. We used thapsigargin, a potent ER stressor that inhibits theER calcium pump, to activate PERK and induce eIF2α phosphorylation.Thapsigargin treatment resulted in a 4.9-fold induction in luciferaseactivity in a 384 well format with a Z factor of 0.5 (FIG. 1B). Thisformat was used to screen 106,281 compounds covering a wide chemicalspace. We identified 460 hits (0.43%) (FIG. 1C), which were furthervalidated in an 8-point dose-response assay using the same reporter. Wefurther triaged the compounds by discarding inhibitors that alsoaffected the IRE1 branch of the UPR using an XBP1-luciferase splicingreporter. Less than half (187 hits) of our initial hits proved unique tothe PERK branch. We next used an orthogonal secondary screen thatemployed a different reporter (bi-cistronic ATF4-dGFPIRES-mCherry)stably integrated into a different cell line (U205 cells). The read-outof this latter screen was microscopy-based, which allowed us tosimultaneously assess acute toxicity by cell counting, further reducingthe number of viable hits to 77. As a tertiary screen, we testedcompounds for their ability to inhibit ER stress-elicited induction ofendogenous ATF4 by Western blot analysis. Twenty-eight compounds passedthis test and were analyzed further.

A symmetric bisglycolamide, ISRIB, is a potent inhibitor of PERKsignaling. One of the 28 compounds was of particular interest because ofits high potency in cells (library compound IC₅₀=40 nM). This compound(henceforth referred to as “ISRIB” for Integrated Stress Responseinhibitor) is a symmetric bis-glycolamide, containing a centralbi-substituted cyclohexane, and can exist as two diastereomers, cis andtrans (FIG. 2A). We synthesized both isomers and tested their ability toinhibit the ATF4-luciferase reporter (FIG. 2B). Trans-ISRIB proved100-fold more potent (IC₅₀=5 nM) than cis-ISRIB (IC₅₀=600 nM),indicating that the compound's interaction with its cellular target isstereospecific. Given the two-order-of-magnitude difference in activityin this assay, the measured activity of cis-ISRIB may be anover-estimate, as we cannot exclude a small contamination withtrans-ISRIB, which is far more potent. The lower IC₅₀ of trans-ISRIBrelative to the compound in the small molecule library indicates thatthe library likely contains a mixture of the two stereoisomers. Allfurther experiments in this study were carried out with the synthesizedtrans-isomer of ISRIB.

ISRIB is PERK-branch specific but does not impair PERK phosphorylation.We next determined at which step ISRIB blocks ATF4 production. To thisend, we first probed the phosphorylation status of PERK by Westernblotting. PERK phosphorylation is indicative of its activation byautophosphorylation and can be recognized by reduced mobility onSDS-polyacrylamide gels. Notably, ISRIB did not inhibit the mobilityshift of PERK observed in ER-stressed cells (FIG. 2C). Rather, weobserved an exaggerated mobility shift, indicative of increasedphosphorylation of PERK upon ER stress, induced by either thapsigarginor tunicamycin (an inhibitor of N-linked glycosylation). In each case,ATF4 and XBP1s were produced upon ER stress induction. In agreement withthe behavior of the reporters described above, ISRIB blocked productionof endogenous ATF4, whereas XBP1 mRNA splicing (FIG. 2D) and XBP1sproduction persisted (FIG. 2C). As shown below (cf. FIG. 5D), ISRIB alsodid not affect the ATF6-branch of the UPR. We conclude that ISRIBspecifically blocks signaling of the PERK-branch of the UPR.

ISRIB-treated cells are resistant to eIF2α phosphorylation. Given thatPERK phosphorylation was not diminished in ISRIB-treated, ER-stressedcells, we next directly assessed eIF2α phosphorylation. We measured thelevels of phosphorylated eIF2α using an antiphospho-eIF2α antibody-basedassay to quantify phosphorylation at serine 51 (see Methods). Uponinduction of ER stress by tunicamycin or thapsigargin, phosphorylationof eIF2α increased over time, reaching a 4- and 7-fold increase after120 minutes respectively (FIG. 3A). Unexpectedly, ISRIB did not blockeIF2α phosphorylation under either of these ER stress-inducingconditions. On the contrary, 120 min after tunicamycin addition, ISRIBfurther increased the level of eIF2α phosphorylation, approaching thatobtained with thapsigargin. ISRIB alone had no effect on eIF2αphosphorylation. These results indicate that ISRIB blocks effectsdownstream of PERK and eIF2α phosphorylation.

One way of explaining why ISRIB blocks ATF4 production yet leaves eIF2αphosphorylation intact is by rendering cells insensitive to the effectsof this phosphorylation event. In agreement with this notion, ISRIBsustained global translation (as monitored by ³⁵S-methionineincorporation into newly synthesized polypeptides) even in the presenceof ER stress (FIG. 3B). After thapsigargin treatment, cells experienceda 40% drop in translation, which was abolished by ISRIB. As predicted bythis result, extracts prepared from mouse embryonic fibroblasts (MEFs)experiencing ER stress showed a pronounced increase in the 80S monosomesat the expense of polyribosomes (FIG. 3C), which was reversed (at leastpartially) by addition of ISRIB. We chose MEFs for this analysis becausethey show stronger translational inhibition in response to ER stressthan HEK293T cells. ISRIB was the only molecule in our collection of 28hits that reversed translational attenuation upon ER-stress.

To further ascertain that cells treated with ISRIB are resistant to theeffects of eIF2α phosphorylation, we transduced an induciblephospho-mimetic allele of eIF2α in which serine 51 was changed to anaspartic acid (S51D) into HEK293T cells. Expression of this allele upondoxycycline addition induced translational attenuation (FIG. 3D) as seenby an increase in the 80S peak and a decrease in the polysomepopulation. ISRIB rescued translation returning it to the levelsobserved in non-induced cells. In conclusion, ISRIB restores translationin cells containing either phospho-eIF2α or eIF2α(S51D), therebyexcluding any pleiotropic effects that might have been caused by thereagents used to activate ER stress.

To rule out that ISRIB exerts non-specific effects on translationindependent of eIF2α phosphorylation, we tested whether ISRIB reverses atranslational block in CAP-mediated initiation. To this end we usedTorin-1, an inhibitor of mTOR that blocks phosphorylation of 4E-BP1 andS6K1, and leads to translational attenuation (17). Addition of Torin-1to MEFs led to an increase in the 80S peak and reduction in the polysomepopulation to a similar degree as shown above in cells treated with ERstressors or expressing eIF2α(S51D) (FIG. 3E, compare with FIGS. 3C and3D). In contrast to these treatments, addition of ISRIB did not reversethe effect of Torin-1 on translation. Therefore, the ability of ISRIB toblock translational attenuation is specific to eIF2α phosphorylation.

If ISRIB makes cells insensitive to eIF2α phosphorylation, it should notmatter which kinase phosphorylates eIF2α. To test this prediction, wesubjected cells to amino acid starvation, which activates the eIF2αkinase GCN2 and leads to ATF4 production. In addition, we used arecently identified small molecule activator to induce eIF2αphosphorylation by activating HRI, another eIF2α kinase (18). Asexpected, ISRIB blocked ATF4 induction after activation of either GCN2or HRI (FIG. 3F). Under both conditions, PERK was not activated as shownby a lack of mobility shift and presence of eIF2α phosphorylation (datanot shown). These data suggest that ISRIB is a bona fide ISR inhibitorthat blocks signaling downstream of all eIF2α kinases.

Both CHOP and GADD34 are transcriptional targets of ATF4. Thus, blockingATF4 accumulation with ISRIB should result in a reduction in thetranscriptional induction of the mRNAs encoding these targets. As shownin FIG. 4A, GADD34 and CHOP mRNAs accumulated in ER-stressed U2OS cells,and ISRIB significantly reduced their induction. In agreement, weobserved no CHOP accumulation after induction of ER stress inISRIB-treated cells (FIG. 4B). Thus ISRIB impairs the transcriptionalnetwork governed by ATF4 during the ISR.

Cell Culture. HEK293T, TREx293, U205, Hela, and mouse embryonicfibroblasts (MEFs) were maintained at 37° C., 5% CO2 in DMEM mediasupplemented with 10% FBS, L-glutamine and antibiotics (penicillin andstreptomycin).

Generation of ATF4 Reporter Constructs and Cell Lines for Small-MoleculeScreening

ATF4 reporters were constructed by fusing the human full-length ATF45′-UTR (NCBI Accession BC022088.2) in front of the firefly luciferase(FLuc) or a destabilized eGFP (dEGFP) coding sequences lacking theinitiator methionine.

The ATF4-FLuc reporter was generated by cloning a PCR-product containingthe ATF4 full-length 5′-UTR (from +1 position a the transcription startsite down to one nucleotide after the terminator codon of the seconduORF) flanked by KpnI/XhoI and BglII sites at the 5′ and 3′ ends,respectively, into the KpnI-BglII sites of pCAX-F-XBP1-Luc. Theresulting construct, pCAX-ATF4-FLuc, was then digested with BamHI,blunted with T4 DNA polymerase, and then digested with XhoI. Theresulting fragment was then subcloned into the retroviral expressionvector pLPCX (Clontech) after digesting it with HindIII, blunting withT4 DNA polymerase and then digesting with XhoI to generatepLPCX-ATF4-FLuc (DAA-312). DAA-312 was used to produce recombinantretroviruses using standard methods and the resulting viral supernatantwas used to transduce HEK293T cells, which were then subsequentlyselected with puromycin to generate a stable cell line employed in theprimary screen.

The ATF4-dEGFP reporter was generated using a PCR fusion-based approach.A PCR product containing the ATF4 full-length 5′ leader sequence (from+1 position a the transcription start site) fused to the eGFP codingsequence 1 nucleotide downstream of the terminator codon of the seconduORF, and flanked by BamHI and EcoRI, was cloned into the cognate sitesof pEGFP-N3 (Clontech) to generate pCMV-ATF4-eGFP. To destabilize theeGFP fusion protein and increase the dynamic range of the reporter,residues 422-461 of mouse ornithine decarboxylase (mODC1), correspondingto its PEST sequence (42), were fused to the C-terminus of the ATF-eGFPfusion protein. To such end, the corresponding mODC1 coding sequence wasamplified by PCR and cloned into the BstXI and EcoRI sites ofpCMV-ATF4-eGFP. The resulting construct was designated pCMV-ATF4-d2EGFP.To further destabilize the ATF4-d1EGFP fusion protein, alaninesubstitutions E428A, E430A, E431A (42) were introduced in the ODC1 PESTsequence to generate pCMV-ATF4-d1EGFP. The ATF4-d1EGFP coding sequencewas then excised from the expression vector using BamHI and EcoRI andsubcloned into the BglII-EcoRI sites of the retroviral expression vectorpLPCX (Clontech) to generate pLPCX-ATF4-d2EGFP. Lastly, a fusion PCRproduct containing the encephalomyocarditis virus internal ribosomalentry site (EMCV-IRES) upstream of the monomeric cherry (mCherry) codingsequence and flanked by EcoRI and NotI recognition sites was subclonedinto the cognate sites of pLPCX-ATF4-d1EGFP, thereby generatingpLPCX-ATF4-d1EGFP-IRES-mCherry (DAA-361). DAA-361 was used to producerecombinant retroviruses using standard methods and the resulting viralsupernatant was used to transduce U2OS cells, which were thensubsequently selected with puromycin to generate a stable cell lineemployed in the secondary screen.

Generation of the inducible eIF2α phosphomimetic mutant construct andcell line

The coding sequences of wild type mouse eIF2α, phosphomimetic (S51D)mutant was amplified by PCR from a mammalian expression vector (kindgift of David Ron). BamHI and EcoRI recognition sites were engineeredinto the primers. In addition a Kozak consensus sequence and aN-terminal FLAG epitope tag were engineered in the forward primer. Theresulting PCR products were subcloned into the cognate sites of thetetracycline-inducible retroviral expression vectorpRetroX-Tight-Pur-GOI (Clontech). 293T target cells stably expressingthe reverse tetracycline transactivator (rtTA) were generated bystandard retroviral transduction using VSV-G pseudotyped retrovirusesencoding rtTA (pRetroX-Tet-On Advanced, Clontech) and selected withGeneticyn. These cells were subsequently transduced with a VSV-Gpseudotyped retrovirus, encoding the eIF2α(S51D) (DAA-A681) mutantallele, resulting in a puromycin-selected, tetracycline inducible,stable cell line.

Generation of the Inducible 6×HIS-3×FLAG-hsATF6-Alpha Cell Line

6×His-3×FLAG-hsATF6-alpha was generated by PCR from p3×FLAGCMV7.1-ATF6(43) and cloned into pcDNA5/FRT/TO.pcDNA5/FRT/T0-6×His-3×FLAG-hsATF6-alpha was co-transfected with pOG44into Flp-In TRex cells (44) according to manufacturers instructions(Invitrogen). After selection with 100 μg/ml Hygromycin B (GoldBiotechnology) single colonies were isolated, expanded and tested forexpression of tagged ATF6.

High-Throughput Primary Screen

HEK293T cells carrying the ATF4 luciferase reporter were plated onpoly-lysine coated 384 well plates (Greiner) at 30,000 cells per well.Cells were treated the next day with 100 nM thapsigargin and 10 μM ofthe library compounds (diversity library of 106,281 compounds) for 6 h.Luminescence was measured using One Glo (Promega) as specified by themanufacturer. The primary screen had a Z′=0.5 and its hit rate was 0.6%(compounds were considered a hit if their luciferase readouts werebeyond three standard deviations of the mean luminescence intensity ofthapsigargin treated cells, which corresponded to 54% inhibition). Ofthese, only 187 compounds did not hit an XBP1-luciferase splicingreporter used as proxy to measure activation of the IRE1 branch of theUPR. Thus, these were considered unique to the PERK branch and werecherry-picked for further analysis.

High-Content Microscopy-Based Secondary Screen

U2OS cells carrying the ATF4-dGFP-IRES-Cherry reporter were plated in 96well plates and treated with 100 nM Thapsigargin and 10 μM of thecherry-picked compounds for 8 h. Cells were stained with Hoechst 33258and were visualized using an automated microscope (InCell Analyzer 2000,GE Healthcare). Data acquisition and image analyses were performed withthe INCell Developer Toolbox Software, version 1.9 (GE Healthcare).Compounds that blocked induction of the ATF4-dGFP reporter, did notblock the accumulation of mCherry downstream of the IRES, and weredeemed nontoxic as determined by cell number measured by countingnuclei, were repurchased for further analyses.

Pharmacokinetics of ISRIB

Intra-peritoneal (ip), and intra-venous (iv) routes of administrationwere performed on 6-7 wk old female CD-1 mice (Harlan Laboratories).Animals received a single, 5 mg/kg dose in groups of threemice/compound/route of administration. For ip and iv dosing ISRIB wasdissolved in DMSO then diluted 1:1 in Super-Refined PEG 400 (Croda).Blood (80 ul) was collected from the saphenous vein at intervalspost-dosing (20 min, 1 h, 3 h, 8 h, 24 h) in EDTA containing collectiontubes (Sarstadt CB300) and plasma was prepared for analysis. Compoundswere detected by time-of-flight mass spectroscopy.

TABLE 1 Pharmacokinetic parameters of ISRIB The data is represented asthe mean (n = 3). Parameters mouse ip dose (mg/kg) 5 AUC (ng*h/ml) 3318F % 13.8 CLt (ml/h) 8.31826 Vd (ml) 96.0261 T½ (h) 8.43

Molecular Action of ISRIB. To date, we have synthesized and assayed morethan 75 analogs, which demonstrate a tractable structure-activityrelationship (to be published elsewhere). The analyses have identifiedsites on the molecule where affinity tags and/or crosslinking moietiescan be added, which promise to aid in target identification. Based onprevious insights on how cells can become resistant to eIF2αphosphorylation, we consider two likely scenarios by which ISRIB couldact:

First, ISRIB could weaken the effects of the non-productive interactionof phospho-eIF2α with eIF2B, thereby increasing the available eIF2α-GEFactivity in the cell, restoring the concentration of ternary complexthat can engage in translation initiation. Precedence for thispossibility derives from genetic studies in S. cerevisiae, where themolecular mechanism of regulation by eIF2α phosphorylation was firstdiscovered. As in mammalian cells, amino acid starvation in yeast leadsto GCN2 activation and eIF2α phosphorylation, resulting in overalltranslational down-regulation and translational induction of atranscriptional activator, GCN4, mediated by uORFs in the 5′UTR of itsmRNA (4). eIF2B is a conserved protein complex comprised of fivedifferent subunits, two of which form the catalytic core, and theremaining three have regulatory roles. Mutations in different eIF2Bsubunits can elicit a phospho-eIF2α resistant phenotype (23-25). Thesemutations have been proposed to act either by weakening the interactionof phospho-eIF2α with eIF2B, reducing its ability to outcompetenon-phosphorylated eIF2 for binding, or by allowing binding ofphospho-eIF2α to the mutant eIF2B in a manner that is conducive tonucleotide exchange. ISRIB could be altering the affinity ofphospho-eIF2α for eIF2B or overcoming the nonproductive interaction thatblocks GTP loading, mimicking the effect of these mutations.

Second, ISRIB could increase the activity of eIF2B, so that the residualamount not engaged with phospho-eIF2α is sufficient to sustain normallevels of ternary complex. Precedence for this possibility derives fromstudies in macrophages, where engagement of toll-like receptor (TLR) 4results in activation of the catalytic activity of eIF2B (26). Thisactivation results from engagement of the TLR-signaling pathway thatinduces a phosphatase removing a constitutively present inhibitoryphosphate from the eIF2B ε-subunit. Pathogens utilize this mechanism tocircumvent translational attenuation and CHOP production under prolongedstress-inducing conditions (27). Similarly, ISRIB could activate orinhibit a signaling pathway that modulates eIF2B activity.

B. Impairment of adaptation to ER stress

As previously shown, cells homozygous for non-phosphorylatable eIF2α,eIF2α(S51A), are unable to cope with ER stress properly, leading toreduced viability (19). This indicates that events downstream of eIF2αphosphorylation are required to resolve the stress. As shown in FIG. 5A,ISRIB treatment of wild-type cells had similar consequences.Importantly, addition of ISRIB alone did not affect cell viability, asjudged by the number of colonies that form after acute treatment. Bycontrast, ISRIB addition caused a strong synergistic effect onER-stressed cells, reducing colony number and size significantly morethan ER-stress alone. This reduction in cell survival resulted fromactivation of apoptosis as the activity of the executioner caspases 3and/or 7 was significantly induced under these conditions (FIG. 5B)(20).

The notion that ER stress remains unmitigated in ISRIB-treated cells issupported by sustained activation of all three UPR sensors. First, asshown in FIG. 2C, PERK was hyper-phosphorylated. Second, cellsexpressing an IRE1-GFP fusion protein showed prolonged foci formation(FIG. 5C), indicative of IRE1 oligomerization. Third, we observedprolonged ER stress-induced proteolytic processing of ATF6 (FIG. 5D).Importantly, in the absence of ER stress ISRIB treatment alone did notinduce any of these sensors (FIGS. 3F and 5C).

Alpha Screen for phospho-S51 eIF2α

U2OS cells were plated on 96 well plates and left to recover overnight.Cells were treated with either with 2 μg/ml tunicamycin or 100 nMthapsigargin in the presence or absence of 100 nM ISRIB or with ISRIBalone for the indicated and the level of eIF2α phosphorylation wasdetermined using the AlphaScreen SureFire eIF2α(p-Ser51) Assay kit(Perkin Elmer) following the manufacturer's recommendations. Plates wereread in an Envision Xcite Multilabel Reader using the standard AlphaScreen settings.

Metabolic Labeling

HEK293T cells were seeded on 12 well plates, allowed to recoverovernight and treated for the indicated times with the indicatedcompounds. The cells were subsequently switched to media lackingmethionine and cysteine supplemented with the indicated compounds and 50μCi of ³⁵S-methionine (Perkin Elmer) for 20 min. Cells were lysed byaddition of SDS-PAGE loading buffer. Lysates were sonicated and equalamounts were loaded on SDS-PAGE gels (BioRad). The gel was dried andradioactive methionine incorporation was detected by exposure to aphosphor-screen and visualized with a Typhoon 9400 Variable Mode Imager(GE Healthcare).

Live Cell Imaging

T-REx293 cells carrying GFP-IRE1 were imaged as described in Li et al,PNAS (45).

Caspase3/7 Activation

Hela cells were plated in 96 well Corning plates at 0.4×10⁴ cells/well24 hours prior to imaging. On the day of experiment, DMEM media wasreplaced with F12 media with appropriate concentration of inhibitors andER stress inducers and caspase 3/7 reagent at 1:1000 dilution (EssenBioscience #4440). Cells were imaged in the IncuCyte FLR live cellimaging system at 2 hour intervals for 70 hours. In order to quantifythe total number of cells, Vybrant DyeCycle Green staining solution (1μM) was added directly to the well immediately after the finalCaspase-3/7 scan and incubated for 1 h prior to acquiring final images.Data was analyzed using IncuCyte analysis software.

qRT-PCR

U2OS cells were plated on 96 well plates and allowed to recoverovernight. Cells were treated for the indicated times with the indicatedcompounds, lysed and cDNA was synthesized using the PowerSYBR GreenCells-to-CT kit (Ambion) following the manufacturer's recommendations.The reactions were ran in an Opticon 2 thermal cycler (BioRad) andanalyzed with the Opticon Monitor v3 software (BioRad). The followingoligonucleotides were used for the amplification reaction: Human GADD34:5′-GTAGCCTGATGGGGTGCTT-3′ (SEQ ID NO:1) and 5′-TGAGGCAGCCGGAGATAC-3′(SEQID NO:2); Human CHOP: 5′-AGCCAAAATCAGAGCTGGAA-3′ (SEQ ID NO:3) and5′-TGGATCAGTCTGGAAAAGCA-3′(SEQ ID NO:4); Human GAPDH:5′-TGGAAGATGGTGATGGGATT-3′ (SEQ ID NO:5) and 5′-AGCCACATCGCTCAGACAC-3′(SEQ ID NO:6).

TAQMAN® Assay to Measure XBP1 mRNA Splicing

cDNA obtained with the POWERSYBR® GREEN CELLS-TO-CT™ kit (Ambion) asdescribed above was used for the TAQMAN® Assay. TAQMAN® assays were setup using IQ™ Supermix (BioRad), 250 nM of each outer primer, 200 nMFAM-XBP1U probe, or 100 nM HEX-XBP1S probe. The reactions were then runon a real-time DNA Engine OPTICON® 2 PCR thermal cycler (BioRad) andanalyzed with the OPTICON MONITOR® v3 software

(BioRad). The outer primers employed for the human XBPlunspliced/spliced(u/s) TAQMAN® assay were: 5′-GAAGCCAAGGGGAATGAAGT-3′ (SEQ ID NO:7), and5′-GAGATGTTCTGGAGGGGTGA-3′ (SEQ ID NO:8). TAQMAN® probes specific forhuman XBP1 s or XBP1u were: 5′-FAM-CAGCACTCAGACTACGTGCACCTCTG-BHQ1-3′(SEQ ID NO:9), and 5′-HEX-TCTGCTGAGTCCGCAGCAGGTGCA-BHQ1-3′ (SEQ IDNO:10). A person of ordinary skill in the art will understand themeaning of the terms “HEX”, “FAM”, and “BHQ-1” as they are used forTAQMAN® probes.

RNA Isolation and Semi-Quantitative RT-PCR

Total RNA from treated or untreated HEK293T cells was extracted usingTRIzol (Invitrogen) following the manufacturer's recommendations. 500 ngof total RNA were reverse transcribed using the SuperScriptVilo cDNASynthesis kit (Invitrogen). The cDNA was diluted 1 in 10 in TE (pH=8)and 1% of the total reaction was used as a template for the PCRamplification reactions. The XBP1 primers flank the 26-nucleotide intronand produce both spliced (222 bp) and unspliced (248 bp) amplicons. ThePCR products were resolved in 2.5% agarose. The followingoligonucleotides were used for the amplification reaction: for humanXBP1, 5′-ACTGGGTCCAAGTTGTCCAG-3′ (SEQ ID NO:11) and5′-GGAGTTAAGACAGCGCTTGG-3′(SEQ ID NO:12); for human GAPDH5′-TGGAAGATGGTGATGGGATT-3′ (SEQ ID NO:13) and5′-AGCCACATCGCTCAGACAC-3′(SEQ ID NO:14).

Protein Analysis

Cells were lysed in SDS-PAGE loading buffer (1% SDS, 62.5 mM Tris-HCl pH6.8, 10% glycerol). Lysates were sonicated and equal amounts were loadedon SDS-PAGE gels (BioRad). Proteins were transferred onto nitrocelluloseand probed with primary antibodies diluted in Tris-buffered salinesupplemented with 0.1% Tween 20 and 5% bovine serum albumin. Thefollowing antibodies were used: CREB-2 (C-20) (1:800) (Santa CruzBiotechnologies); PERK (D11A8) (1:1000), PERK (C33E10) (1:1000), eIF2α(9722) (1:1000), phospho-eIF2α (Ser51) (D9G8) XP (3398) (1:1000) (CellSignaling Technology); XBP1s (C-terminus) (1:500) (BioLegend); M2 Flag(1:1000) (Sigma). An HRP-conjugated secondary antibody (Amersham) wasemployed to detect immune-reactive bands using enhancedchemiluminescence (SuperSignal, Thermo Scientific).

Immunofluorescence

U2OS cells were seeded on Slide Flasks (Thermo Scientific) 18 h prior toprocessing for immunofluorescence. Cells (60% confluent) were fixed with4% paraformaldehyde in PBS for 15 min. The cells were then rinsed 3times with PBS and permeabilized with 0.3% Triton X-100. The fixed cellswere rinsed 3 times with PBS and blocked for 1 h at room temperature inPBS supplemented with 0.1% Triton X-100 and 5% normal goat serum. Thecells were then incubated overnight at 4° C. with an anti-CHOP mousemonoclonal antibody (Cell Signaling Technology L63F7) at a 1:1000dilution in blocking buffer. The next morning the slides were washed 3times (5 min each time) with PBST (PBS-0.1% Triton X-100). The slideswere then incubated for 1 h at room temperature in a 1:500 dilution (inblocking buffer) of secondary anti-mouse antibody labeled with Alexa Dye488 (Molecular Probes). The slides were then washed 3 additional timeswith PBST. The cells were then counterstained with rhodaminephalloidin(1:1,000 in PBS) for 10 min at room temperature to reveal the actincytoskeleton. Lastly, the slides were mounted using Vectashield (Vector)mounting medium and imaged using a Zeiss Axiovert 200M epifluorescencemicroscope.

Polysome Gradients

Mouse Embryonic Fibroblasts (MEFs) or TREx-293 cells expressing eIF2α(S51D) were seeded on 150 mm plates and allowed to grow to 80%confluence. Cells were then induced with 25 nM doxycycline for 14 h andsubsequently treated with the appropriate compounds for the indicatedtimes. 100 μg/ml of cycloheximide was added to the cells for 1 minbefore lysis. Cells were washed twice with PBS supplemented with 100μg/ml cycloheximide and subsequently lysed in 20 mM Tris pH 7.4, 200 mMNaCl, 15 mM MgCl, 1 mM DTT, 8% Glycerol, 100 μg/ml cycloheximide, 1%Triton X-100 and EDTA-free protease inhibitor tablets (Roche). Cellswere scraped, collected, triturated with a 25^(7/8) gauge needle, andthe homogenate was centrifuged for 10 min at 10,000×g. The supernatantwas loaded on a 10-50% sucrose gradient and sedimented in a SW40 rotorat 150,000×g for 2.4 h. The gradients were fractionated using a pistongradient fractionator (BioComp) and UV absorbance at 254 nm wasmonitored using a UV-Monitor (BioRad).

ISRIB can influence cell fate. As a signaling network withinterconnected signaling branches, the UPR exhibits both cytoprotectiveand pro-apoptotic functions. When faced with ER stress, PERK-mediatedtranslational attenuation contributes to adaptation by reducing the loadof newly synthesized proteins that are translocated into the ER (13). Inaddition, induction of the transcription regulator ATF4 upregulates manygenes that increase the protein folding capacity in the ER. Both ofthese activities serve to reestablish homeostasis, balancing the proteinfolding load and protein folding capacity in the ER lumen. Thisreasoning is supported by the increased sensitivity to ER stressexhibited by MEFs that lack PERK or ATF4, as well as MEFs that carry anon-phosphorylatable knock-in allele of eIF2α(S51A) (13,19,28). Inagreement, we show that ISRIB decreases the viability of cells that aresubjected to ER-stress. In these cells, ISRIB sustains IRE1 and ATF6activation, indicating that ER stress remains unmitigated in the absenceof PERK signaling. As some cancer cells sustain an activated UPR to aidin their survival, ISRIB could provide a new therapeutic approach tocancer chemotherapy. In agreement, a PERK-specific inhibitordemonstrates antitumor activity in a human pancreatic tumor xenograftmodel (29). The deleterious synergistic effect between ER-stress andISRIB may be generally advantageous to kill cancer cells, especiallythose derived from secretory lineages that have increased secretory loadand increased basal levels of ER stress (including myelomas, andpancreatic and breast cancers).

Importantly, by acting downstream of eIF2α phosphorylation, ISRIB blocksmultiple stress effectors (i.e., all eIF2α kinases). During tumorgrowth, hypoxic conditions and a lack of nutrients can activate bothPERK and GCN2, and PERK or GCN2 MEFs give rise to significantly smallertumors in mouse xenograft models than their wild-type counterparts(30,31). Hence both kinases have pro-survival roles in tumordevelopment. By blocking signaling by both kinases, ISRIB displaysunique properties that may be beneficial in reducing cellular fitness oftumor cells.\

Results in a multiple myeloma xenograft model suggest that ISRIB hasantitumor activity in subcutaneous plasmacytomas arising from RPMI 8226cells (FIG. 22). In this experiment, ISRIB, given orally to tumorbearing mice, resulted in a significant reduction in the rate of tumorgrowth. At the endpoint of the study, there was a 50% reduction in tumorsize in animals dosed with ISRIB.

C. Memory Studies

Reversing translational attenuation with ISRIB synergistically reducesthe viability of cells subjected to PERK-activation by chronicendoplasmic reticulum (ER) stress. eIF2α phosphorylation has beenimplicated in memory consolidation by modulating new protein synthesisin the brain. Remarkably, wild-type mice injected with ISRIB displaysignificant enhancement in both spatial and fear-associated learning.These results show that memory consolidation in normal animals isinherently limited by the ISR and that ISRIB can release this break. Assuch, ISRIB promises to contribute to our understanding and treatment ofcognitive disorders.

Eight to ten-week-old male C57BL/6J mice were used for behavioralexperiments. Food and water were provided ad libitum, and mice were kepton a 12:12 h light/dark cycle (lights on at 08:00 h). All procedurescomplied with Canadian Council on Animal Care guidelines

Morris Water Maze

Mice were trained in a water pool of 100 cm diameter with a hiddenplatform of 10 cm diameter. Mice were handled daily for 3 days beforethe experiment, and the training protocol consisted of 1 swimming trialper day. Each mouse swam until it found the hidden platform or 120 s,when it was gently guided to the platform and stayed there for 10 sbefore being returned to the cage. Immediately after the swimming trialthe mice were injected intraperitoneally with ISRIB (0.25 mg/kg insaline, 1% DMSO). For the probe test, the platform was removed and eachmouse was allowed to swim for 60 s, while its swimming trajectory wasmonitored with a video tracking system (HVS Image, Buckingham).

Contextual Fear Conditioning

Mice were handled for 3 days and thereafter injected dailyintraperitoneally with ISRIB (0.25 mg/kg in saline, 1% DMSO) for 4consecutive days. One hour after the last injection the mice weretrained with the protocol, which consisted of a 2-min period of contextexploration, followed by a single foot shock of 0.35 mA for 1 s. Themice were returned to their home cage 1 min after the shock. One and 24h after training, the mice were tested for contextual fear memory byplacing the animals in the conditioning context for a 4-min period. Theincidence of freezing was scored in 5-s intervals as either “freezing”or “not freezing”. Percent of freezing indicates the number of intervalsin which freezing was observed divided by total number of 5-s intervals.Statistical analyses were done by Student's t tests and one-way ANOVAfollowed by between-group comparisons using Tukey's posthoc test.

Cannulation and Auditory Fear Conditioning

Male Sprague Dawley rats (275-350 g) were used for cannulation asdescribed in Migues et al, 2010 (46). ISRIB (0.05 mg/ml, 5 μl) wasinfused bilaterally into the amygdala immediately after auditory fearconditioning training. The infusion was performed with a microinjector(28 gauge) connected to a Hamilton syringe with plastic tubing at a rateof 0.4 μl/min. To allow for the solution containing ISRIB to diffusefrom the tip of the cannula into the tissue, the microinjector stayed inthe cannula for one additional minute. Training protocol for auditoryfear conditioning consisted of a 2-min period of context A exploration,followed by one pairing of a tone (2800 Hz, 85 dB, 30 s) with aco-terminating foot shock (0.75 mA, 1 s). Rats were returned to theirhome cage 1 min after the shock. Test for auditory fear memory consistedof a 2 min acclimatizing period to the context B (pre-CS), followed bytone presentation (CS) (2800 Hz, 85 dB, 30 s). Freezing time wasmeasured and percent of freezing was calculated. At the end of theexperiment, cannula placement was checked by examining 50 μm brainsections stained with formal-thionin under a light microscope.

ISRIB increases long-term memory in rodents. eIF2α(+/S51A) heterozygotemice display enhanced memory, while induction of the eIF2α kinase PKR inbrain pyramidal cells impairs memory (21,22). Based on theseobservations, we wondered whether treatment of mice with ISRIB wouldaffect memory. ISRIB showed favorable properties in pharmacokineticprofiling experiments indicating sufficient bioavailability for in vivostudies (Table 1). To explore ISRIB's effects on memory, we injectedmice intraperitoneally with ISRIB and tested hippocampus-dependentspatial learning. To this end, we trained mice in a Morris water maze,in which animals learn to associate visual cues with the location of asubmerged hidden platform. Because we were looking for memoryenhancement, we used a weak training protocol. As shown in FIG. 6A,ISRIB-treated mice reached the hidden platform significantly faster(escape latency after 5 days of training=16.4+/−4.8 s) compared tovehicle treated controls (68.1+/−20 s, p<0.05). The difference wasalready pronounced by days 3 and 4. In agreement with these results,ISRIB-treated mice significantly preferred the target quadrant in a“probe test” conducted at the end of the training sessions, in which theplatform was removed from the pool (p<0.05; FIG. 6B) and showedincreased crossing of the platform location (p<0.05; FIG. 6C).

We next tested contextual fear conditioning, which represents adifferent kind of hippocampus-dependent learning in which eIF2αphosphorylation has also been implicated to play a role (22). In theseexperiments, we paired a particular environmental context (a differentcage) with a foot shock. In this case the context acts as the“conditioned stimulus, CS” and is associated with the foot shock, the“unconditioned stimulus, US”. ISRIB-treated mice showed increasedfreezing upon presentation of the conditioned environment 24 h aftertraining as compared to vehicle treated mice (p<0.05; FIG. 6D). Nodifferences were observed in short-term memory (1 h) between these twotreatments. Taken together, we conclude that treatment with ISRIBenhances both hippocampus-dependent spatial learning andhippocampus-dependent contextual fear conditioning.

To test learning associated with a different region of the brain, weexplored the effects of ISRIB on auditory fear conditioning, whichdepends on the amygdala. In this type of learning a tone (CS) is pairedwith a foot shock (US). In these experiments, we injected ISRIB orvehicle directly into the amygdala of rats via cannulation.ISRIB-treated rats showed a significant increase over vehicle-injectedrats in the level of freezing when presented with the tone (CS) at 24 h(long-term memory, p<0.05; FIG. 6E). By contrast, we observed nodifference between ISRIB- and vehicle-treated rats at 3 h (short-termmemory). As expected, both ISRIB- and vehicle-treated rats showedsimilar freezing responses prior to training (pre-CS). Taken together,these data suggest that long-term memory is selectively enhanced inISRIB-treated animals.

ISRIB and brain function. The importance of eIF2/eIF2B function in thehuman brain is underscored by familial diseases caused by mutations inthese factors. One example is Childhood Ataxia with CNS Hypomyelination(CACH), also known as Vanishing White Matter disease (VWM), which hasbeen mapped to mutations in different subunits of eIF2B (32). A secondexample links a familial intellectual disability syndrome to a mutationin the γ-subunit of eIF2 complex (33).

Several lines of genetic evidence in mice suggest thatphosphorylation-dependent regulation of eIF2α phosphorylation is acritical hub for the control of synaptic plasticity (as assessed by latelong-term potentiation (L-LTP) in brain slices) and memory consolidation(as assessed in behavioral tasks in animals). In particular, thethreshold for induction of L-LTP is reduced and memory consolidation isenhanced in mice lacking GCN2 or PKR and in mice heterozygous fornon-phosphorylatable eIF2α(S51A), which have reduced levels of eIF2αphosphorylation (22,34,35). As we show here, ISRIB pharmacologicallyphenocopies these genetic manipulations in behavioral tasks by renderingcells insensitive to eIF2α phosphorylation. In agreement, treatment ofmice with a PKR inhibitor was reported to enhance memory consolidation,and, conversely, treatment with salubrinal, an inhibitor that prolongseIF2α phosphorylation, to block L-LTP and memory consolidation (22,35).

eIF2α phosphorylation results in a dual effect on gene expression: aglobal translational diminution and translational upregulation of selectmRNA, including ATF4 mRNA. Both may be important to explain the observedeffects on L-LTP and memory. It has long been appreciated that newprotein synthesis is required for memory consolidation and that ATF4represses CREB-mediated transcription of “memory genes” (36,37). Indeed,this latter function of ATF4 in memory consolidation is evolutionarilyconserved from Aplysia to rodents (38-40). Because a small physiologicalincrease in the level of eIF2α phosphorylation that does notsignificantly alter overall translation is sufficient to induce ATF4,production of this transcription factor can be finely tuned in neuronalcells by perhaps selective activation of different eIF2αkinases. Theobserved effects of ISRIB may therefore result from overcoming effectscaused by a relatively small level of regulatory phosphorylation that isdistinct from the high level resulting from ER stress-inducing agents.In light of this reasoning, a therapeutic window may exist in whichISRIB's effects as memory enhancer can be exploited without encounteringlong-term toxic consequences.

ISRIB increases memory consolidation, allowing pharmacologicalenhancement of the brain's ability to learn. Evolution therefore did notarrive at a maximally optimized process, imposing a break (via eIF2αphosphorylation) on memory consolidation. This mechanism may underscorethe importance of filtering memories before committing them to long-termstorage. Indeed, eIF2α phosphorylation also plays a role in dynamicrestructuring of memory, as indicated by studies showing that ablationof PERK in the brain impairs behavioral flexibility (41). Our findingsraise the possibility that ISRIB or compounds with related activitiescould serve as invaluable tools in deciphering these higher order brainfunctions and perhaps be beneficial as a therapeutic agent effectingmemory improvement in diseases associated with memory impairment.

D. Synthesis of ISRIB

General Methods

Commercially-available reagents and solvents were used as received.Silica gel chromatography was performed using a Biotage Isolera Fourflash purification system with Silicycle SiliaSep™ cartridges. ¹H NMRspectra were recorded on a Varian INOVA-400 400 MHz spectrometer.Chemical shifts are reported in δ units (ppm) relative to residualsolvent peak. Coupling constants (J) are reported in hertz (Hz). LCMSanalyses were carried out using a Waters 2795 separations moduleequipped with a Waters 2996 photodiode array detector, a Waters 2424 ELSdetector, a Waters micromass ZQ single quadropole mass detector, and an)(Bridge C18 column (5 μm, 4.6×50 mm).

Synthesis of Bisglycolamides

trans-ISRIB:2-(4-Chlorophenoxy)-N-[(1r,4r)-4-[2-(4-chlorophenoxy)acetamido]cyclohexyl]acetamide

To a mixture of (1r,4r)-cyclohexane-1,4-diamine (20 mg, 0.18 mmol) intetrahydrofuran:water (1:1, 1 ml) were sequentially added potassiumcarbonate (73 mg, 0.53 mmol) and 4-chlorophenoxyacetyl chloride (56 μl,0.36 mmol). Upon addition of the acid chloride, a white solidimmediately formed. The reaction mixture was vigorously stirred atambient temperature for 30 min. Water (2.5 ml) was added. The mixturewas vigorously vortexed then centrifuged, and the water was decanted.This washing protocol was repeated with potassium bisulfate (1% aq, 2.5ml), water (2.5 ml), and diethyl ether (2×2.5 ml). The resulting wetwhite solid was dried by partially dissolving indichloromethane/methanol (10/1, 10 ml) and gravity filtering through anAutochem 4.5 mL reaction tube. The residual undissolved product wasextracted from the wet filter cake by adding dichloromethane (4×4.5 ml)and gravity filtering. The combined filtrate was concentrated usingrotary evaporation to afford 51 mg (65%) of the title compound as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ 7.91 (d, J=8.1 Hz, 2H), 7.31(d, J=9.0 Hz, 4H), 6.94 (d, J=9.0 Hz, 4H), 4.42 (s, 4H), 3.55 (br. s.,2H), 1.73 (br. d, J=5.9 Hz, 4H), 1.30 (quin, J=10.5 Hz, 4H); LC-MS:m/z=451 [M+H, ³⁵Cl×2]+, 453 [M+H, ³⁵Cl, ³⁷Cl]⁺.

cis-ISRIB: 2-(4-chlorophenoxy)-N-[(1s,4s)-4-[2-(4-chlorophenoxy)acetamido]cyclohexyl]acetamide

To a mixture of (1s,4s)-cyclohexane-1,4-diamine (21 μl, 20 mg, 0.18mmol) in tetrahydrofuran:water (1:1, 1 ml) were sequentially addedpotassium carbonate (73 mg, 0.53 mmol) and 4-chlorophenoxyacetylchloride (56 μL, 0.36 mmol). The reaction mixture was vigorously stirredat ambient temperature for 1.5 h then partitioned between 30 mL of 1:1dichloromethane:KHSO₄ (10% aq.). After separating the organic layer, itwas sequentially washed with water (1×10 ml) and brine (1×10 ml) thendried by gravity filtration using an Autochem 4.5 mL reaction tube. Thefiltrate was concentrated and loaded onto a Silicycle 4 g SiO₂ columnusing a minimal amount of dichloromethane (˜2 ml). The product waseluted with acetone in dichloromethane (0%-50%). Product-containingfractions were combined and concentrated to afford 56 mg (71%) of thetitle compound as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 7.76 (d,J=7.0 Hz, 2H), 7.32 (d, J=9.0 Hz, 4H), 6.94 (d, J=9.0 Hz, 4H), 4.47 (s,4H), 3.70 (br. s., 2H), 1.44-1.67 (m, 8H); LC-MS: m/z=451 [M+H,₃₅Cl×2]₊, 453 [M+H, ³⁵Cl, ³⁷Cl]⁺.

E. Increasing Protein Expression

Compounds described herein (e.g. ISRIB) can be used to increase proteinproduction (e.g. antibodies) in cell systems such as antibody-producingcells or hybridomas by increasing overall protein translation. ISRIB canalso be used to increase recombinant protein production in cell-freesystems (rabbit reticulocyte or Hela in vitro translation system). ISRIBblocks the effects of eIF2α phosphorylation on translation initiationand thus enhances overall translation in cells where thisphosphorylation event normally imposes a brake. We showed that inmultiple myeloma cells, which produce and secrete antibodies, additionof ISRIB leads to increased translation both in ER-stressed andun-stressed cells (FIGS. 23A and 23B, respectively). By increasingoverall translation, addition of ISRIB may allow for increased antibodyproduction. ISRIB enhances translation of an exogenously added mRNA in arabbit-reticulocyte in vitro translation system (FIG. 24).

TABLE 2 IC50 SMDC (nM) ID Cell-luc Smiles 751591   250O═C(N[C@@H]1CC[C@@H](NC(COC2═CC═C(OC)C═C2)═O)CC1)COC3═CC═C(Cl)C═C3751592   >10000O═C(N[C@@H]1CC[C@@H](NC(COC2═CC═C(OCC3═CC═CC═C3)C═C2)═O)CC1)COC4═CC═C(Cl)C═C4751593   384O═C(COC1═CC═C(C═C1)Cl)N[C@@H]2CC[C@H](CC2)NC(COC3═CC═CC═C3)═O 751594  69O═C(N[C@@H]1CC[C@@H](NC(COC2═C(Cl)C═C(Cl)C═C2)═O)CC1)COC3═CC═C(Cl)C═C3751595   >10000O═C(N[C@@H]1CC[C@@H](NC(CC2═CC═C(C)C═C2)═O)CC1)COC3═CC═C(Cl)C═C3751596   >10000O═C(N[C@@H]1CC[C@@H](NC(C2═CC═C(Cl)C═C2)═O)CC1)COC3═CC═C(Cl)C═C3751597   >10000O═C(N[C@@H]1CC[C@@H](NC(COC2═CC═C(O)C═C2)═O)CC1)COC3═CC═C(Cl)C═C3754123   >10000COc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(OC)cc3)cc1754124   >10000Clc1ccc(OCC(═O)N[C@@H]2CC[C@@H](CC2)NC(═O)COc3ccc(Cl)cc3Cl)cc1 754128  95 Cc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1754125   >10000Clc1ccc(CCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)CCc3ccc(Cl)cc3)cc1 754127   13FC(F)(F)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C(F)(F)F)cc1754126   327 Cc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(C)cc3)cc1755854.1 2700Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)N(CC#C)C(═O)COc3ccc(Cl)cc3)cc1755854.4 64Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)N(CC#C)C(═O)COc3ccc(Cl)cc3)cc1755854.2 440Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)N(CC#C)C(═O)COc3ccc(Cl)cc3)cc1755855   142 Clc1ccc(OCC(═O)N[C@@H]2C[C@H](C2)NC(═O)COc3ccc(Cl)cc3)cc1755856   1000 Clc1ccc(OCC(═O)N[C@@H]2C[C@@H](C2)NC(═O)COc3ccc(Cl)cc3)cc1757095   >10000O═C(COc1ccc(cc1)C#N)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C#N757096   >10000CS(═O)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)S(═O)C)cc1757131   270 Fc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(F)cc3)cc1757130   >10000[O−][N+](═O)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)[N+](═O)[O−])cc1757132   3000Clc1ccc(OCC(N[C@H]2CC[C@](NC(COc3ccc(Cl)cc3)═O)(C#C)CC2)═O)cc1 755854.3100 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)N(CC#C)C(═O)COc3ccc(Cl)cc3)cc1750213.2 3 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1835195   170 Clc1ccc(OCCN[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1835196   1400Clc1ccc(OCCN(CCOc2ccc(Cl)cc2)[C@@H]3CC[C@H](CC3)NC(═O)COc4ccc(Cl)cc4)cc1835197   >10000Clc1ccc(OCC(═N)N[C@@H]2CC[C@H](CC2)N(Cc3cn(CCOCCOCCOCCNC(═O)CCCC[C@@H]4SCC5NC(═O)NC45)nn3)C(═O)COc6ccc(Cl)cc6)cc1757257   48 Fc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1757258   10FC(F)(F)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1757259   263Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C#N)cc1757260   >10000CS(═O)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1757261   >10000Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)CSc3ccc(Cl)cc3)cc1757262   >10000Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)CS(═O)c3ccc(Cl)cc3)cc1757263   >10000Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)CS(═O)(═O)c3ccc(Cl)cc3)cc1757264.1 100 Clc1ccc(OCC(═N)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1757264.2 34 Clc1ccc(OCC(═N)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1835087   2700Clc1ccc(OCC(═O)N[C@@H]2CC[C@](CC2)(NC(═O)COc3ccc(Cl)cc3)c4cn(CCOCCOCCOCCNC(═O)CCCC[C@H]5SCC6NC(═O)NC56)nn4)cc1835089   700 Clc1ccc(NCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1757149   37Clc1ccc(OCC(═O)N[C@@H]2CC[C@](CC2)(NC(═O)COc3ccc(Cl)cc3)C#C)cc1843983   >10000CC(═O)c1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1 843984  1000 CC(═O)c1cccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)c1843987   75[O−][N+](c1c(OCC(N[C@H]2CC[C@H](NC(COc3ccc(Cl)cc3)═O)CC2)═O)ccc(Cl)c1)═O811769   >10000 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)CSc3ccccc3)cc1811770   900 Fc1ccc(SCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1811771   >10000Clc1ccc(OCC(═O)N[C@@H]2C[C@H](C2)NC(═O)CSc3ccc(Cl)cc3)cc1811772   >10000 Fc1ccc(SCC(═O)N[C@@H]2C[C@H](C2)NC(═O)COc3ccc(Cl)cc3)cc1811773   >10000 Clc1ccc(OCC(═O)N[C@@H]2C[C@H](C2)NC(═O)CSc3ccccc3)cc1873882   >10000 Clc1ccc(OCC(═O)NCc2cccc(NC(═O)COc3ccc(Cl)cc3)c2)cc1873883   2600C[C@@H]1NC(═O)N[C@@H]1CCCCCC(═O)NCCOCCOCCOCCn2cc(nn2)[C@@]3(CC[C@@H](CC3)NC(═O)COc4ccc(Cl)cc4)NC(═O)COc5ccc(Cl)cc5873972   >10000O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═C(C(C)═O)C═C(Cl)C═C3)═O)CC2873973   >10000O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═C(C(C4═CC═CC═C4)═O)C═C(Cl)C═C3)═O)CC2757264-L3 60O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C═C3)═N)CC2874796   >10000O═C(COC1═CC═C(Cl)C═C1)NCC2═CC═C(CNC(COC3═CC═C(Cl)C═C3)═O)C═C2 874797  122 O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(OCC3═CC═C(Cl)C═C3)═O)CC2874798   200O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(C(C)C)C═C3)═O)CC2874799   210O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(C(C)OC3═CC═C(Cl)C═C3)═O)CC2874800   2O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(Cl)═C3)═O)CC2874801   2.4O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(C)═C3)═O)CC2874802   2O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(F)═C3)═O)CC2874803   4O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(C(F)(F)F)═C3)═O)CC2874804   3O═C(COC1═CC═C(Cl)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C([N+]([O−])═O)═C3)═O)CC2874805   >10000 ClC(C═CC═C1)═C1CNC[C@H]2CC[C@H](CNCC3═CC═CC═C3Cl)CC2885253   >10000O═C(CC1═CC═C(Cl)C═C1)NCC2═CC═C(CNC(CC3═CC═C(Cl)C═C3)═O)C═C2885254   >10000O═C(OC1═CC═C(Cl)C═C1)NCC2═CC═C(CNC(OC3═CC═C(Cl)C═C3)═O)C═C2 885255   30O═C(OC1═CC═C(Cl)C═C1)NC[C@H]2CC[C@H](NC(COC3═CC═C(Cl)C═C3)═O)CC2885256   152O═C(CC1═CC═C(Cl)C═C1)NC[C@H]2CC[C@H](NC(COC3═CC═C(Cl)C═C3)═O)CC2885257   >10000O═C(OCC1═CC═C(Cl)C═C1)N[C@@H](CC2)CC[C@H]2NC(OCC3═CC═C(Cl)C═C3)═O102509   53 ClC(C═C1)═CC═C1OCC(NC2═CC═C(NC(COC3═CC═C(Cl)C═C3)═O)C═C2)═O912562   125 Fc1cccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)c1912563   334 Fc1cccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)c1F912564   220Fc1cc(F)cc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)c1912565   >10000Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3cc(Cl)cc(Cl)c3)cc1 912566  149 CC(═O)c1cc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)ccc1Cl913815   >10000Clc1ccc(OC(═O)NC[C@@H]2CC[C@@H](CNC(═O)Oc3ccc(Cl)cc3)CC2)cc1 914582  5.2 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)c(l)c3)cc1914583   12C[Si](C)(C)C#Cc1cc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)ccc1Cl914584   12Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)c(c3)C#C)cc1914989   5.6 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(l)cc3)cc1Clc1ccc(OCC(N[C@H]2CC[C@@H](NC(COc3ccc(Cl)cc3)═O)CC2)═O)cc1Clc1ccc(OCC(NC2CCC(NC(COc3ccc(Cl)cc3)═O)CC2)═O)cc1 916348   160C[Si](C)(C)C#Cc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(Cl)cc3)cc1916353   14 Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C#C)cc1916727   520C[C@@H]1NC(═O)N[C@@H]1CCCCCC(═O)NCCOCCOCCOCCn2cc(nn2)c3ccc(OCC(═O)N[C@@H]4CC[CH](CC4)NC(═O)COc5ccc(Cl)cc5)cc3916728   1,270C[C@@H]1NC(═O)N[C@@H]1CCCCCC(═O)NCCOCCOCCOCCn2cc(nn2)c3cc(OCC(═O)N[C@@H]4CC[C@H](CC4)NC(═O)COc5ccc(Cl)cc5)ccc3Cl916744   >10000FC(F)(F)C1(NN1)c2ccc(OCC(═O)N[C@@H]3CC[C@H](CC3)NC(═O)COc4ccc(l)cc4)cc2916751   >10000O═C(COC1═CC═C(C#C)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(C4(N═N4)C(F)(F)F)C═C3)═O)CC2916784   51 ldccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(l)cc3)cc1916785   >10000C[Si](C)(C)C#Cc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C#C[Si](C)(C)C)cc1916786   158O═C(COc1ccc(cc1)C#C)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)C#C955278   >10000O═C(COC1═CC═C(C#C)C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(C4(N═N4)C(F)(F)F)═C3)═O)CC2916846   200Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)c4cn(CCCCCC(═O)OCCCCCNC(═O)CCCC[C@H]5SC[C@@H]6NC(═O)N[C@H]56)nn4)cc1916847   900Clc1ccc(OCC(═O)N[C@@H]2CC[C@H](CC2)NC(═O)COc3ccc(cc3)c4cn(CCCCCC(═O)OCCCCCNC(═O)CCCC[C@H]5SC[C@@H]6NC(═O)N[C@H]56)nn4)cc1957866   232O═C(COC1═CC═C(N═[N+]═[N−])C═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(C#C)C═C3)═O)CC2957885   0.8O═C(COC1═CC═C(Cl)C(Cl)═C1)N[C@@H]2CC[C@@Hl(NC(COC3═CC═C(Cl)C(Cl)═C3)═O)CC2957886   3O═C(COC1═CC═C(Cl)C(C)═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(C)═C3)═O)CC2957887   11O═C(COC1═CC═C(Cl)C(C(F)(F)F)═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(C(F)(F)F)═C3)═O)CC2957888   0.6O═C(COC1═CC═C(Cl)C(F)═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C(F)═C3)═O)CC2957889   11 O═C(COC1═CC═C(Cl)C([N+]([O−])═O)═C1)N[C@@H]2CC[C@@H](NC(COC3═CC═C(Cl)C([N+]([O−])═O)═C3)═O)CC2957914   40O═C(COC1═CC(Cl)═C(Cl)C═C1)N[C@H]2CC[C@H](NC(COC3═CC═C(Cl)C(Cl)═C3)═[NH2+])CC2.[Cl−]957915   344O═C(COC1═CC(F)═C(Cl)C═C1)N[C@H]2CC[C@H](NC(COC3═CC═C(Cl)C(F)═C3)═[NH2+])CC2957916   122O═C(COC1═CC(F)═C(Cl)C═C1)N[C@H]2CC[C@H](NC(COC3═CC═C(Cl)C(F)═C3)═[NH2+])CC2SMDC ID Structure 751591  

751592  

751593  

751594  

751595  

751596  

751597  

754123  

754124  

754128  

754125  

754127  

754126  

755854.1

755854.4

755854.2

755855  

755856  

757095  

757096  

757131  

757130  

757132  

755854.3

750213.2

835195  

835196  

835197  

757257  

757258  

757259  

757260  

757261  

757262  

757263  

757264.1

757264.2

835087  

835089  

757149  

843983  

843984  

843987  

811769  

811770  

811771  

811772  

811773  

873882  

873883  

873972  

873973  

757264-L3

874796  

874797  

874798  

874799  

874800  

874801  

874802  

874803  

874804  

874805  

885253  

885254  

885255  

885256  

885257  

102509  

912562  

912563  

912564  

912565  

912566  

913815  

914582  

914583  

914584  

914989  

916348  

916353  

916727  

916728  

916744  

916751  

916784  

916785  

916786  

955278  

916846  

916847  

957866  

957885  

957886  

957887  

957888  

957889  

957914  

957915  

957916  

References 1. Wek R C, Jiang H-Y, Anthony TG. Coping with stress: eIF2kinases and translational control. Biochem. Soc. Trans. 2006 February;34(Pt 1):7-11. 2. Hinnebusch A G, Lorsch J R. The mechanism ofeukaryotic translation initiation: new insights and challenges. ColdSpring Harb Perspect Biol. 2012; 4(10). 3. Krishnamoorthy T, Pavitt G D,Zhang F, Dever T E, Hinnebusch A G. Tight binding of the phosphorylatedalpha subunit of initiation factor 2 (eIF2alpha) to the regulatorysubunits of guanine nucleotide exchange factor eIF2B is required forinhibition of translation initiation. Mol Cell Biol. 2001 August;21(15):5018-30. 4. Hinnebusch A G. Translational regulation of GCN4 andthe general amino acid control of yeast. Annu. Rev. Microbiol. 2005;59:407-50. 5. Jackson R J, Hellen CUT, Pestova T V. The mechanism ofeukaryotic translation initiation and principles of its regulation. NatRev Mol Cell Biol. 2010 Feb. 1; 11(2):113-27. 6. Harding H P, Novoa I,Zhang Y, Zeng H, Wek R, Schapira M, et al. Regulated translationinitiation controls stress-induced gene expression in mammalian cells.Mol Cell. 2000 November; 6(5):1099-108. 7. Palam L R, Baird T D, Wek RC. Phosphorylation of eIF2 facilitates ribosomal bypass of an inhibitoryupstream ORF to enhance CHOP translation. Journal of BiologicalChemistry. 2011 Apr. 1; 286(13):10939-49. 8. Vattem K M, Wek R C.Reinitiation involving upstream ORFs regulates ATF4 mRNA translation inmammalian cells. Proc Natl Acad Sci USA. 2004 Aug. 3; 101(31):11269-74.9. Ma Y, Brewer J W, Diehl J A, Hendershot L M. Two distinct stresssignaling pathways converge upon the CHOP promoter during the mammalianunfolded protein response. J. Mol. Biol. 2002 May 17; 318(5):1351-65.10. Pavitt G D, Ron D. New insights into translational regulation in theendoplasmic reticulum unfolded protein response. Cold Spring HarbPerspect Biol. 2012 June; 4(6). 11. Ron D, Walter P. Signal integrationin the endoplasmic reticulum unfolded protein response. Nat Rev Mol CellBiol. 2007 July; 8(7):519-29. 12. Gardner B M, Walter P. Unfoldedproteins are Ire1-activating ligands that directly induce the unfoldedprotein response. Science. 2011 Sep. 30; 333(6051):1891-4. 13. Harding HP, Zhang Y, Bertolotti A, Zeng H, Ron D. Perk is essential fortranslational regulation and cell survival during the unfolded proteinresponse. Mol Cell. 2000 May; 5(5):897-904. 14. Walter P, Ron D. Theunfolded protein response: from stress pathway to homeostaticregulation. Science. 2011 Nov. 25; 334(6059):1081-6. 15. Tabas I, Ron D.Integrating the mechanisms of apoptosis induced by endoplasmic reticulumstress. Nat Cell Biol. 2011 Mar. 1; 13(3):184-90. 16. Shore GCG, PapaFRF, Oakes SAS. Signaling cell death from the endoplasmic reticulumstress response. Current Opinion in Cell Biology. 2011 Apr. 1;23(2):143-9. 17. Thoreen C C, Chantranupong L, Keys H R, Wang T, Gray NS, Sabatini D M. A unifying model for mTORC1-mediated regulation of mRNAtranslation. Nature. 2012 May 3; 485(7396):109-13. 18. Chen T, Ozel D,Qiao Y, Harbinski F, Chen L, Denoyelle S, et al. Chemical geneticsidentify eIF2α kinase heme-regulated inhibitor as an anticancer target.Nature Chemical Biology. 2011 Sep. 1; 7(9):610-6. 19. Lu P D, Jousse C,Marciniak S J, Zhang Y, Novoa I, Scheuner D, et al. Cytoprotection bypre-emptive conditional phosphorylation of translation initiation factor2. EMBO J. 2004 Jan. 14; 23(1):169-79. 20. Salvesen G S, Ashkenazi A.Snapshot: caspases. Cell. 2011 Oct. 14; 147(2):476-476.el. 21. Jiang Z,Belforte J E, Lu Y, Yabe Y, Pickel J, Smith C B, et al. eIF2alphaPhosphorylation-dependent translation in CA1 pyramidal cells impairshippocampal memory consolidation without affecting general translation.Journal of Neuroscience. 2010 Feb. 17; 30(7):2582-94. 22. Costa-MattioliM, Gobert D, Stern E, Gamache K, Colina R, Cuello C, et al. eIF2alphaphosphorylation bidirectionally regulates the switch from short- tolong-term synaptic plasticity and memory. Cell. 2007 Apr. 6;129(1):195-206. 23. Vazquez de Aldana C R, Hinnebusch A G. Mutations inthe GCD7 subunit of yeast guanine nucleotide exchange factor eIF-2Bovercome the inhibitory effects of phosphorylated eIF-2 on translationinitiation. Mol Cell Biol. 1994 May; 14(5):3208-22. 24. Pavitt G D,Ramaiah K V, Kimball S R, Hinnebusch A G. eIF2 independently binds twodistinct eIF2B subcomplexes that catalyze and regulateguanine-nucleotide exchange. Genes Dev. 1998 Feb. 15; 12(4):514-26. 25.Pavitt G D, Yang W, Hinnebusch A G. Homologous segments in threesubunits of the guanine nucleotide exchange factor eIF2B mediatetranslational regulation by phosphorylation of eIF2. Mol Cell Biol. 1997March; 17(3):1298-313. 26. Woo C W, Kutzler L, Kimball S R, Tabas I.Toll-like receptor activation suppresses ER stress factor CHOP andtranslation inhibition through activation of eIF2B. Nat Cell Biol. 2012Feb. 1; 14(2):192-200. 27. Woo C W, Cui D, Arellano J, Dorweiler B,Harding H, Fitzgerald K A, et al. Adaptive suppression of the ATF4-CHOPbranch of the unfolded protein response by toll-like receptorsignalling. Nat Cell Biol. 2009 December; 11(12):1473-80. 28. Harding HP, Zhang Y, Zeng H, Novoa I, Lu P D, Calfon M, et al. An integratedstress response regulates amino acid metabolism and resistance tooxidative stress. Mol Cell. 2003 March; 11(3):619-33. 29. Axten J M,Medina J R, Feng Y, Shu A, Romeril S P, Grant S W, et al. Discovery of7-methyl-5-(1-{[3-(trifluoromethyl)phenyl]acetyl}-2,3-dihydro-1H-indol-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(GSK2606414), a potent and selective first-in-class inhibitor of proteinkinase R (PKR)-like endoplasmic reticulum kinase (PERK). J Med Chem.2012 Aug. 23; 55(16):7193-207. 30. Bi M, Naczki C, Koritzinsky M, FelsD, Blais J, Hu N, et al. ER stress-regulated translation increasestolerance to extreme hypoxia and promotes tumor growth. EMBO J. 2005Oct. 5; 24(19):3470-81. 31. Ye J, Kumanova M, Hart L S, Sloane K, ZhangH, De Panis D N, et al. The GCN2-ATF4 pathway is critical for tumourcell survival and proliferation in response to nutrient deprivation.EMBO J. 2010 Jun. 16; 29(12):2082-96. 32. Li W, Wang X, Van Der Knaap MS, Proud C G. Mutations Linked to Leukoencephalopathy with VanishingWhite Matter Impair the Function of the Eukaryotic Initiation Factor 2BComplex in Diverse Ways. Mol Cell Biol. 2004 Apr. 15; 24(8):3295-306.33. Borck G, Shin B-S, Stiller B, Mimouni-Bloch A, Thiele H, Kim J-R, etal. eIF2γ Mutation that Disrupts eIF2 Complex Integrity LinksIntellectual Disability to Impaired Translation Initiation. Mol Cell.2012 Oct. 34. Costa-Mattioli M, Gobert D, Harding H, Herdy B, Azzi M,Bruno M, et al. Translational control of hippocampal synaptic plasticityand memory by the eIF2alpha kinase GCN2. Nature. 2005 Aug. 25;436(7054):1166-73. 35. Zhu P J, Huang W, Kalikulov D, Yoo J W, Placzek AN, Stoica L, et al. Suppression of PKR Promotes Network Excitability andEnhanced Cognition by Interferon-γ-Mediated Disinhibition. Cell. 2011Dec. 9; 147(6):13-3. 36. Sutton M A, Schuman E M. Dendritic proteinsynthesis, synaptic plasticity, and memory. Cell. 2006 October 6;127(1):49-58. 37. Klann E, Dever T E. Biochemical mechanisms fortranslational regulation in synaptic plasticity. Nat. Rev. Neurosci.2004 December; 5(12):931-42. 38. Bartsch D, Ghirardi M, Skehel P A, KarlK A, Herder S P, Chen M, et al. Aplysia CREB2 represses long-termfacilitation: relief of repression converts transient facilitation intolong-term functional and structural change. Cell. 1995 Dec. 15;83(6):979-92. 39. Yin J C, Wallach J S, Del Vecchio M, Wilder E L, ZhouH, Quinn W G, et al. Induction of a dominant negative CREB transgenespecifically blocks long-term memory in Drosophila. Cell. 1994 Oct. 7;79(1):49-58. 40. Chen A, Muzzio I A, Malleret G, Bartsch D, Verbitsky M,Pavlidis P, et al. Inducible enhancement of memory storage and synapticplasticity in transgenic mice expressing an inhibitor of ATF4 (CREB-2)and C/EBP proteins. Neuron. 2003 Aug. 14; 39(4):655-69. 41. Trinh MAM,Kaphzan H H, Wek RCR, Pierre P P, Cavener DRD, Klann E E. Brain-specificdisruption of the eIF2α kinase PERK decreases ATF4 expression andimpairs behavioral flexibility. Cell Rep. 2012 Jun. 28; 1(6):676-88. 42.Li X, Zhao X, Fang Y, Jiang X, Duong T, Fan C, et al. Generation ofdestabilized green fluorescent protein as a transcription reporter. JBiol Chem. 1998 Dec. 25; 273(52):34970-5. 43. Shen J, Chen X, HendershotL, Prywes R. ER stress regulation of ATF6 localization by dissociationof BiP/GRP78 binding and unmasking of Golgi localization signals.Developmental Cell. 2002 July; 3(1):99-111. 44. Cohen H R, Panning B.XIST RNA exhibits nuclear retention and exhibits reduced associationwith the export factor TAP/NXF1. Chromosoma. 2007 August; 116(4):373-83.45. Li H, Korennykh A V, Behrman S L, Walter P. Mammalian endoplasmicreticulum stress sensor IRE1 signals by dynamic clustering. Proceedingsof the National Academy of Sciences. 2010 Sep. 14; 107(37):16113-8. 46.Migues P V, Hardt O, Wu D C, Gamache K, Sacktor T C, Wang Y T, et al.PKMzeta maintains memories by regulating GluR2-dependent AMPA receptortrafficking. Nat. Neurosci. 2010 May; 13(5):630-4. 47. Bi M, Naczki C,Koritzinsky M, Fels D, Blais J, Hu N, Harking H, Novoa I, Varia M,Raleigh J, Scheuner D, Kaufman R J, Bell J, Ron D, Wouters B G, KoumenisC. 2005. ER stress-regulated translation increases tolerance to extremehypoxia and promotes tumor growth. EMBO J. 24:3470-3481. 48.Bobrovnikova-Marjon E, Pytel D, Vaites L P, Singh N, Koretzky G A, DiehlJ A. 2010. PERK promotes cancer cell proliferation and tumor growth bylimiting oxidative DNA damage. Oncogene 29: 3881-3895. 49.Avivar-Valderas A, Bobrovnikova-Marjon E, Diehl A, Nagi C, Debnath J,Aguirre-Guiso J A 2011. PERK integrates autophagy and oxidative stressresponses to promote survival during extracellular matrix detachment.Mol Cel Biol 31: 3616-3629. 50. Axten J M., Medina J. R., Feng Y., ShuA., Romeril S. P. et al. 2012. Discovery of7-methy-5(1-{[3-(trifluoromethyl)phenyl]acetyl}-2,3-dihydro-1H-indol-5yl)-7H-pyrrolo[2,3-d]pyrimidin-4 amine (GSK2606414), a potent and selective first-inclass inhibitor of protein kinase R (PKR)-like endplasmic reticulumkinase (PERK). J. Med. Chem. 55(16): 7193-7207 51. Ye J. Kumanova M.,Hart L. S., Sloane K., Zhang H. et al. 2010. The GCN2-ATF4 pathway iscritical for tumour cell survival and proliferation in response tonutrient deprivation. EMBO J. 29: 2082-2096. 52. Moreno J A, Radford H,Peretti D, Steinert J R, Verity N, Martin M G, Halliday M, Morgan J,Dinsdale D, Ortori C A, Barrett D A, Tsaytler P, Bertolotti A, Willis AE, Bushell M, Mallucci G R. 2012. Sustained translational repression byeIF2α-P mediates prion neurodegeneration. Nature 485:507-511. 53. PavittG D and Proud C G. 2009. Protein synthesis and its control in neuronalcells with a focus on vanishing white matter disease. Biochem Soc Trans37:1298-1310. 54. Costa-Mattioli M. Gobert D., Harding H., Herdy B. AzziM., Bruno M. et al, 2005. Translational control of hippocampal synapticplasticity and memory by the eIF2α kinase GCN2. Nature 436:1166-1173.55. Costa-Mattioli M., Gobert D., Stern E., Garnache K., Colina Rl,Cuello C., Sossin W., Kaufman R., Pelletier J., Rosenblum et al. 2007.eIF2α phosphorylation bidirectionally regulates the switch from short tolong term synaptic plasticity and memory. Cell 129: 195-206. 56. Zhu P.J, Huan W., Kalikulov D., Yoo J. W., Placzek A. N., Stoica L, Zhou H.,Bell J. C., Frielander M. J., Krnjevic K., Noebels J. L., Costa-MattioliM. 2011. Suppression of PKR promotes network excitability and enhancedcognition by interferon-γ-mediated disinhibition. Cell 147: 1384-1396.57. Borck G., Shin B. S., Stiller B., et al 2012. eIF2γ mutation thatdisrupts eIF2 complex integrity links intellectual disability toimpaired translation initiation. Mol Cell 48:1-6. 58. Zeenko V. V., WangC, Majumder M, Komar A. A., Snider M. D., Merrick W. C., Kaufman R. J.and Hatzoglou M. (2008). An efficient in vitro translation system frommammalian cell lacking translational inhibition caused by eIF2phosphorylation. RNA 14: 593-602. 59. Mikami S., Masutani M., SonenberN., Yokoyama S. and Imataka H. 2006. An efficient mammalian cell-freetranslation system supplemented with translation factors. Protein Expr.Purif 46: 348-357.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

What is claimed is:
 1. A method of improving memory consolidation and long term memory in a patient, said method comprising administering a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, to said patient, wherein said compound has the formula:

wherein L² and L⁴ are independently, —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen, halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃, —C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCF₃, —OCHF₂, —N₃, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R² and R⁴ are independently ═NR⁷, ═O, or ═S; R⁸ and R⁹ are independently hydrogen, halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; b and d are independently 0 or 1; and z5 and z6 are independently an integer from 0 to
 5. 2. The method of claim 1, wherein the compound has the formula:

wherein: R^(5.1) and R^(6.1) are independently hydrogen, halogen, —CF₃, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl, substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted or unsubstituted 5 to 6 membered heteroaryl,

R^(5.2) and R^(6.2) are independently hydrogen, halogen, —CCSi(CH₃)₃, —CF₃, —NO₂, —CN, —N₃, substituted or unsubstituted C₁-C₄ alkyl, substituted or unsubstituted 2 to 4 membered heteroalkyl, substituted or unsubstituted 5 to 6membered heteroaryl,


3. The method of claim 2, wherein, R^(5.1) and R^(6.1) are independently —Cl, —I, —CF₃, —CH₃, or —CCH; and R^(5.2) and R^(6.2) are independently hydrogen, —Cl, —F, —I, —CCSi(CH₃)₃, —CF₃, —NO₂, —CH₃, or —CCH.
 4. The method of claim 1, wherein the compound is


5. The method of claim 1, wherein the compound is


6. The method of claim 1, wherein the patient has traumatic brain injury.
 7. A method of treating vanishing white matter disease in a patient, said method comprising administering a therapeutically effective amount of a compound, or a pharmaceutically acceptable salt thereof, to said patient, wherein said compound has the formula:

wherein L² and L⁴ are independently —O—, —S—, —S(O)—, —S(O)₂—, substituted or unsubstituted alkylene or substituted or unsubstituted heteroalkylene; R¹, R³, R⁵, R⁶ and R⁷ are independently hydrogen, halogen, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —C(NN)CF₃, —C(NH—NH)CF₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)OH, —NHOH, —OCF₃, —OCHF₂, —N₃, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; R² and R⁴ are independently ═NR⁷, ═O, or ═S; R⁸ and R⁹ are independently hydrogen, halogen, —OCH₃, —OCH₃, —OCH₂Ph, —C(O)Ph, —CH₃, —CF₃, —CCl₃, —CN, —S(O)CH₃, —OH, —NH₂, —COOH, —CONH₂, —NO₂, —C(O)CH₃, —CH(CH₃)₂, —CCSi(CH₃)₃, —CCH, —CH₂CCH, —SH, —SO₂Cl, —SO₃H, —SO₄H, —SO₂NH₂, —NHNH₂, —ONH₂, —NHC═(O)NHNH₂, —NHC═(O)NH₂, —NHSO₂H, —NHC═(O)H, —NHC(O)—OH, —NHOH, —OCF₃, —OCHF₂, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; and b and d are independently 0 or 1; and z5 and z6 are independently an integer from 0 to
 5. 